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A rapid and transient ROS generation by cadmium triggers apoptosis via caspase-dependent pathway in HepG2 cells and this is inhibited through N-acetylcysteine-mediated catalase upregulation
Although reactive oxygen species (ROS) have been implicated in cadmium (Cd)-induced hepatotoxicity, the role of ROS in this pathway remains unclear. Therefore, we attempted to determine the molecular mechanisms relevant to Cd-induced cell death in HepG2 cells. Cd was found to induce apoptosis in the HepG2 cells in a time- and dose-dependent fashion, as confirmed by DNA fragmentation analysis and TUNEL staining. In the early stages, both rapid and transient ROS generation triggered apoptosis via Fas activation and subsequent caspase-8-dependent Bid cleavage, as well as by calpain-mediated mitochondrial Bax cleavage. The timing of Bid activation was coincided with the timing at which the mitochondrial transmembrane potential (MMP) collapsed as well as the cytochrome c (Cyt c) released into the cytosol. Furthermore, mitochondrial permeability transition (MPT) pore inhibitors, such as cyclosporin A (CsA) and bongkrekic acid (BA), did not block Cd-induced ROS generation, MMP collapse and Cyt c release. N-acetylcysteine (NAC) pretreatment resulted in the complete inhibition of the Cd-induced apoptosis via catalase upregulation and subsequent Fas downregulation. NAC treatment also completely blocked the Cd-induced intracellular ROS generation, MMP collapse and Cyt c release, indicating that Cd-induced mitochondrial dysfunction may be regulated indirectly by ROS-mediated signaling pathway. Taken together, a rapid and transient ROS generation by Cd triggers apoptosis via caspase-dependent pathway and subsequent mitochondrial pathway. NAC inhibits Cd-induced apoptosis through the blocking of ROS generation as well as the catalase upregulation.
Oh, Seon-Hee [Research Center for Resistant Cells, College of Medicine, Chosun University, Seosuk-dong, Dong-gu, Gwangju 501-759 (Korea, Republic of); Lim, Sung-Chul [Research Center for Resistant Cells, College of Medicine, Chosun University, Seosuk-dong, Dong-gu, Gwangju 501-759 (Korea, Republic of) and Department of Pathology, College of Medicine, Chosun University, Seosuk-dong, Dong-gu, Gwangju 501-759 (Korea, Republic of)]. E-mail: email@example.com
Catalase Production by Lactobacilli
LACTOBACILLI are considered to be catalase-negative1, or occasionally to produce very small amounts of catalase2. The lactobacilli isolated from dairy products have always been found to be catalase-negative3,4. During recent work on the microflora of Cheddar cheese, we have encountered strains of lactobacilli which possess both strong and weak catalase activity. Three strains of Lactobacillus plantarum, originally isolated by Sherwood3
J. C. Dacre
NSDL National Science Digital Library
The catalase test is used to detect the presence of the enzyme catalase in bacteria. Catalase serves to neutralize the bactericidal effects of hydrogen peroxide. Its concentration in bacteria has been correlated with pathogenicity. This enzymatic test is essential in the scheme of identification for gram-positive organisms and certain gram-negative organisms. It is a primary test used in the differentiation of staphylococci and streptococci.
American Society For Microbiology;
Inhibitors of Catalase Reaction
IT is well known that the activity of catalase is greatly inhibited by very small concentrations of potassium cyanide, hydrogen sulphide and especially hydroxylamine. To these reagents we can now add sodium azide (NaN3) which also acts as a strong inhibitor of catalase.
D. Keilin; E. F. Hartree
Non-heme manganese catalase - the 'other' catalase
Non-heme manganese catalases are widely distributed over microbial life and represent an environmentally important alternative to heme-containing catalases in antioxidant defense. Manganese catalases contain a binuclear manganese complex as their catalytic active site rather than a heme, and cycle between Mn2(II,II) and Mn2(III,III) states during turnover. X-ray crystallography has revealed the key structural elements of the binuclear manganese active site complex that can serve as the starting point for computational studies on the protein. Four manganese catalase enzymes have been isolated and characterized, and the enzyme appears to have a broad phylogenetic distribution including both bacteria and archae. More than 100 manganese catalase genes have been annotated in genomic databases, although the assignment of many of these putative manganese catalases needs to be experimentally verified. Iron limitation, exposure to low levels of peroxide stress, thermostability and cyanide resistance may provide the biological and environmental context for the occurrence of manganese catalases.
Whittaker, James W.
High Dietary Fat Selectively Increases Catalase Expression within Cardiac Mitochondria*
Obesity is a predictor of diabetes and cardiovascular disease. One consequence of obesity is dyslipidemia characterized by high blood triglycerides. It has been proposed that oxidative stress, driven by utilization of lipids for energy, contributes to these diseases. The effects of oxidative stress are mitigated by an endogenous antioxidant enzyme network, but little is known about its response to high fat utilization. Our experiments used a multiplexed quantitative proteomics method to measure antioxidant enzyme expression in heart tissue in a mouse model of diet-induced obesity. This experiment showed a rapid and specific up-regulation of catalase protein, with subsequent assays showing increases in activity and mRNA. Catalase, traditionally considered a peroxisomal protein, was found to be present in cardiac mitochondria and significantly increased in content and activity during high fat feeding. These data, coupled with the fact that fatty acid oxidation enhances mitochondrial H2O2 production, suggest that a localized catalase increase is needed to consume excessive mitochondrial H2O2 produced by increased fat metabolism. To determine whether the catalase-specific response is a common feature of physiological conditions that increase blood triglycerides and fatty acid oxidation, we measured changes in antioxidant expression in fasted versus fed mice. Indeed, a similar specific catalase increase was observed in mice fasted for 24 h. Our findings suggest a fundamental metabolic process in which catalase expression is regulated to prevent damage while preserving an H2O2-mediated sensing of diet composition that appropriately adjusts insulin sensitivity in the short term as needed to prioritize lipid metabolism for complete utilization.
Rindler, Paul M.; Plafker, Scott M.; Szweda, Luke I.; Kinter, Michael
Molecular evolution of maize catalases and their relationship to other eukaryotic and prokaryotic catalases
We have compared the nucleotide and protein sequences of the three maize catalase genes with other plant catalases to reconstruct\\u000a the evolutionary relationship among these catalases. These sequences were also compared with other eukaryotic and prokaryotic\\u000a catalases. Phylogenies based on distances and parsimony analysis show that all plant catalases derive from a common ancestral\\u000a catalase gene and can be divided
Lingqiang Guan; John G. Scandalios
Catalase and glutathione peroxidase mimics
Overproduction of the reactive oxygen species (ROS) superoxide (O2?) and hydrogen peroxide (H2O2) are increasingly implicated in human disease and aging. ROS are also being explored as important modulating agents in a number of cell signaling pathways. Earlier work has focused on development of small catalytic scavengers of O2?, commonly referred to as superoxide dismutase (SOD) mimetics. Many of these compounds also have substantial abilities to catalytically scavenge H2O2 and peroxynitrite (ONOO?). Peroxides have been increasingly shown to disrupt cell signaling cascades associated with excessive inflammation associated with a wide variety of human diseases. Early studies with enzymatic scavengers like SOD frequently reported little or no beneficial effect in biologic models unless SOD was combined with catalase or a peroxidase. Increasing attention has been devoted to developing catalase or peroxidase mimetics as a way to treat overt inflammation associated with the pathophysiology of many human disorders. This review will focus on recent development of catalytic scavengers of peroxides and their potential use as therapeutic agents for pulmonary, cardiovascular, neurodegenerative and inflammatory disorders.
Day, Brian J.
Enantioselective inhibition of dichlorprop on catalase.
The enantioselectivity interaction of 2,4-dichlorprop (DCPP) and catalase were studied, and it was further evaluated with the presence of humus. Both of rac-DCPP and R-DCPP can inhibit the activity of catalase with the concentrations of 0.05-80 mg L(-1), the inhibitory type of rac-DCPP was uncompetitive, and of R-DCPP was complex. The presence of humic acid has changed the inhibitory ability of DCPP on catalase, the inhibition of rac-DCPP disappeared and the inhibition type of R-DCPP mainly became uncompetitive. These results suggest that inhibition of chiral DCPP on catalase is enantioselective. PMID:22961377
Ma, Yun; Jiang, Jihong; Xu, Chao; Lu, Xianting
Tubular Flow Reactor Studies of Immobilized Catalase and Glucose Oxidase.
National Technical Information Service (NTIS)
Catalase was immobilized on porous glass and several nickel silica alumina supports. Catalase immobilized on nickel silica alumina was studied in a tubular plug flow reactor. Several physical mixtures of singly immobilized catalase on nickel silica alumin...
A. D. Traher
21 CFR 173.135 - Catalase derived from Micrococcus lysodeikticus.
Code of Federal Regulations, 2010 CFR
...2012-04-01 2012-04-01 false Catalase derived from Micrococcus lysodeikticus...Preparations and Microorganisms § 173.135 Catalase derived from Micrococcus lysodeikticus. Bacterial catalase derived from Micrococcus...
21 CFR 173.135 - Catalase derived from Micrococcus lysodeikticus.
Code of Federal Regulations, 2010 CFR
...2009-04-01 false Catalase derived from Micrococcus lysodeikticus. 173.135 Section... § 173.135 Catalase derived from Micrococcus lysodeikticus. Bacterial catalase derived from Micrococcus lysodeikticus by a pure...
21 CFR 173.135 - Catalase derived from Micrococcus lysodeikticus.
Code of Federal Regulations, 2010 CFR
...2009-04-01 true Catalase derived from Micrococcus lysodeikticus. 173.135 Section... § 173.135 Catalase derived from Micrococcus lysodeikticus. Bacterial catalase derived from Micrococcus lysodeikticus by a pure...
Catalase negative mutants of Helicobacter pylori.
Nine strains of Helicobacter pylori have been isolated exhibiting spontaneous mutations with a loss of catalase activity. Growth characteristics in vitro were unaffected by the mutation showing that catalase is not essential for growth of Helicobacter pylori. Parent strains and mutants could not be distinguished morphologically from each other when compared by electron microscopy. Restriction endonuclease digestion with HindIII, separated in an 0.7% agarose gel in TBE buffer, showed each pair to be highly related to each other. SDS-PAGE separation of proteins from four mutants and parent strains showed that all mutants lacked a 57 kDa protein. The partial N-terminal sequence of this protein shows homology with maize catalase and may be the subunit of the Helicobacter pylori catalase tetramer. It is concluded that catalase negative mutants of Helicobacter pylori occur spontaneously in vitro, but have not yet been observed in vivo. The paucity of such catalase negative strains in clinical specimens could mean that catalase is a virulence factor in vivo that puts mutants at a selective disadvantage. PMID:1526235
Westblom, T U; Phadnis, S; Langenberg, W; Yoneda, K; Madan, E; Midkiff, B R
Evolution of catalases from bacteria to humans.
Excessive hydrogen peroxide is harmful for almost all cell components, so its rapid and efficient removal is of essential importance for aerobically living organisms. Conversely, hydrogen peroxide acts as a second messenger in signal-transduction pathways. H(2)O(2) is degraded by peroxidases and catalases, the latter being able both to reduce H(2)O(2) to water and to oxidize it to molecular oxygen. Nature has evolved three protein families that are able to catalyze this dismutation at reasonable rates. Two of the protein families are heme enzymes: typical catalases and catalase-peroxidases. Typical catalases comprise the most abundant group found in Eubacteria, Archaeabacteria, Protista, Fungi, Plantae, and Animalia, whereas catalase-peroxidases are not found in plants and animals and exhibit both catalatic and peroxidatic activities. The third group is a minor bacterial protein family with a dimanganese active site called manganese catalases. Although catalyzing the same reaction (2 H(2)O(2)--> 2 H(2)O+ O(2)), the three groups differ significantly in their overall and active-site architecture and the mechanism of reaction. Here, we present an overview of the distribution, phylogeny, structure, and function of these enzymes. Additionally, we report about their physiologic role, response to oxidative stress, and about diseases related to catalase deficiency in humans. PMID:18498226
Zamocky, Marcel; Furtm�ller, Paul G; Obinger, Christian
Evolution of Catalases from Bacteria to Humans
Excessive hydrogen peroxide is harmful for almost all cell components, so its rapid and efficient removal is of essential importance for aerobically living organisms. Conversely, hydrogen peroxide acts as a second messenger in signal-transduction pathways. H2O2 is degraded by peroxidases and catalases, the latter being able both to reduce H2O2 to water and to oxidize it to molecular oxygen. Nature has evolved three protein families that are able to catalyze this dismutation at reasonable rates. Two of the protein families are heme enzymes: typical catalases and catalase�peroxidases. Typical catalases comprise the most abundant group found in Eubacteria, Archaeabacteria, Protista, Fungi, Plantae, and Animalia, whereas catalase�peroxidases are not found in plants and animals and exhibit both catalatic and peroxidatic activities. The third group is a minor bacterial protein family with a dimanganese active site called manganese catalases. Although catalyzing the same reaction (2 H2O2 ? 2 H2O + O2), the three groups differ significantly in their overall and active-site architecture and the mechanism of reaction. Here, we present an overview of the distribution, phylogeny, structure, and function of these enzymes. Additionally, we report about their physiologic role, response to oxidative stress, and about diseases related to catalase deficiency in humans.
Zamocky, Marcel; Furtmuller, Paul G.; Obinger, Christian
Characterization of Catalase from Psychrotolerant Psychrobacter piscatorii T-3 Exhibiting High Catalase Activity.
A psychrotolerant bacterium, strain T-3 (identified as Psychrobacter piscatorii), that exhibited an extraordinarily high catalase activity was isolated from the drain pool of a plant that uses H(2)O(2) as a bleaching agent. Its cell extract exhibited a catalase activity (19,700 U�mg protein(-1)) that was higher than that of Micrococcus luteus used for industrial catalase production. Catalase was approximately 10% of the total proteins in the cell extract of the strain. The catalase (PktA) was purified homogeneously by only two purification steps, anion exchange and hydrophobic chromatographies. The purified catalase exhibited higher catalytic efficiency and higher sensitivity of activity at high temperatures than M. luteus catalase. The deduced amino acid sequence showed the highest homology with catalase of Psycrobacter cryohalolentis, a psychrotolelant bacterium obtained from Siberian permafrost. These findings suggest that the characteristics of the PktA molecule reflected the taxonomic relationship of the isolate as well as the environmental conditions (low temperatures and high concentrations of H(2)O(2)) under which the bacterium survives. Strain T-3 efficiently produces a catalase (PktA) at a higher rate than Exiguobacterium oxidotolerans, which produces a very strong activity of catalase (EktA) at a moderate rate, in order to adapt to high concentration of H(2)O(2). PMID:22408420
Kimoto, Hideyuki; Yoshimune, Kazuaki; Matsuyma, Hidetoshi; Yumoto, Isao
Catalase: A repertoire of unusual features
Catalases are antioxidant enzymes which catalyze the breakdown of hydrogen peroxide to water and oxygen, and are one of the\\u000a oldest enzymes to be studied biochemically. The first crystal structure of a catalase appeared in the year 1980 and it revealed\\u000a the tetrameric nature of the enzyme and presence of channels accessing the deeply buried active site heme. An interesting
Prashen Chelikani; T. Ramana; T. M. Radhakrishnan
A molecular switch and electronic circuit modulate catalase activity in catalase-peroxidases
The catalase reaction of catalase-peroxidases involves catalase-specific features built into a peroxidase core. An arginine, 20 � from the active-site heme, acts as a molecular switch moving between two conformations, one that activates heme oxidation and one that activates oxoferryl heme reduction by H2O2, facilitating the catalatic pathway in a peroxidase. The influence of the arginine is imparted to the
Xavier Carpena; Ben Wiseman; Taweewat Deemagarn; Rahul Singh; Jacek Switala; Anabella Ivancich; Ignacio Fita; Peter C Loewen
Detection of catalase in rat heart mitochondria.
The presence of heme-containing catalase in rat heart mitochondria (20 +/- 5 units/mg) was demonstrated by biochemical and immunocytochemical analysis. Intact rat heart mitochondria efficiently consumed exogenously added H2O2. The rate of H2O2 consumption was not influenced by succinate, glutamate/malate, or N-ethylmaleimide but was significantly inhibited by cyanide. Hydrogen peroxide decomposition by mitochondria yielded molecular oxygen in a 2:1 stoichiometry, consistent with a catalytic mechanism. Mitochondrial fractionation studies and quantitative electron microscopic immunocytochemistry revealed that most catalase was matrix-associated. Electrophoretic analysis and Western blotting of the mitochondrial matrix fraction indicated the presence of a protein with similar electrophoretic mobility to bovine and rat liver catalase and immunoreactive to anti-catalase antibody. Myocardial tissue has a lower catalase-specific activity and a greater mitochondrial H2O2 production/g of tissue than most organs. Thus catalase, representing 0.025% of heart mitochondrial protein, is important for detoxifying mitochondrial derived H2O2 and represents a key antioxidant defense mechanism for myocardial tissue. PMID:1657986
Radi, R; Turrens, J F; Chang, L Y; Bush, K M; Crapo, J D; Freeman, B A
Catalase and NO CATALASE ACTIVITY1 promote autophagy-dependent cell death in Arabidopsis.
Programmed cell death often depends on generation of reactive oxygen species, which can be detoxified by antioxidative enzymes, including catalases. We previously isolated catalase-deficient mutants (cat2) in a screen for resistance to hydroxyurea-induced cell death. Here, we identify an Arabidopsis thaliana hydroxyurea-resistant autophagy mutant, atg2, which also shows reduced sensitivity to cell death triggered by the bacterial effector avrRpm1. To test if catalase deficiency likewise affected both hydroxyurea and avrRpm1 sensitivity, we selected mutants with extremely low catalase activities and showed that they carried mutations in a gene that we named NO CATALASE ACTIVITY1 (NCA1). nca1 mutants showed severely reduced activities of all three catalase isoforms in Arabidopsis, and loss of NCA1 function led to strong suppression of RPM1-triggered cell death. Basal and starvation-induced autophagy appeared normal in the nca1 and cat2 mutants. By contrast, autophagic degradation induced by avrRpm1 challenge was compromised, indicating that catalase acted upstream of immunity-triggered autophagy. The direct interaction of catalase with reactive oxygen species could allow catalase to act as a molecular link between reactive oxygen species and the promotion of autophagy-dependent cell death. PMID:24285797
Hackenberg, Thomas; Juul, Trine; Auzina, Aija; Gwizdz, Sonia; Malolepszy, Anna; Van Der Kelen, Katrien; Dam, Svend; Bressendorff, Simon; Lorentzen, Andrea; Roepstorff, Peter; Lehmann Nielsen, K�re; J�rgensen, Jan-Elo; Hofius, Daniel; Breusegem, Frank Van; Petersen, Morten; Andersen, Stig Uggerh�j
Increased myocardial catalase in rats fed ethanol.
The effects of chronic intake of dietary ethanol upon catalase, an enzyme capable of metabolizing ethanol, as well as upon myocardial morphology and hemodynamics, were studied in the rat. Ethanol, comprising 36% of dietary calories, administered to rats for 5 weeks, was associated with increased myocardial catalase of 45.9 +/- 3.7 IU/mg protein, compared to 21.0 +/- 1.8 IU/mg protein in pair-fed controls. The enzyme activity remained significantly elevated after 18 weeks of ethanol. Hepatic catalase did not differ in these groups. Parallel cytochemical studies confirmed the increase in myocardial catalase by demonstrating an increase in peroxisomes. Gross and light-microscopic examinations revealed no abnormalities at either 5 or 18 weeks. Remarkably few ultrastructural abnormalities were seen in this material fixed by vascular perfusion. Hemodynamic studies after 5 weeks of ethanol revealed decreased left ventricle systolic pressure and decreased mean arterial pressure but no change in ventricular filling pressure. The possibility of catalase playing a metabolic and potentially protective role in rat myocardium chronically exposed to ethanol is discussed. Images Figure 3 Figure 4-6 Figures 1 and 2 Figures 7 and 8 p[389]-a
Fahimi, H. D.; Kino, M.; Hicks, L.; Thorp, K. A.; Abelman, W. H.
Protection of Bacillus pumilus spores by catalases.
Bacillus pumilus SAFR-032, isolated at spacecraft assembly facilities of the National Aeronautics and Space Administration Jet Propulsion Laboratory, is difficult to kill by the sterilization method of choice, which uses liquid or vapor hydrogen peroxide. We identified two manganese catalases, YjqC and BPUM_1305, in spore protein extracts of several B. pumilus strains by using PAGE and mass spectrometric analyses. While the BPUM_1305 catalase was present in six of the B. pumilus strains tested, YjqC was not detected in ATCC 7061 and BG-B79. Furthermore, both catalases were localized in the spore coat layer along with laccase and superoxide dismutase. Although the initial catalase activity in ATCC 7061 spores was higher, it was less stable over time than the SAFR-032 enzyme. We propose that synergistic activity of YjqC and BPUM_1305, along with other coat oxidoreductases, contributes to the enhanced resistance of B. pumilus spores to hydrogen peroxide. We observed that the product of the catalase reaction, gaseous oxygen, forms expanding vesicles on the spore surface, affecting the mechanical integrity of the coat layer, resulting in aggregation of the spores. The accumulation of oxygen gas and aggregations may play a crucial role in limiting further exposure of Bacilli spore surfaces to hydrogen peroxide or other toxic chemicals when water is present. PMID:22752169
Checinska, Aleksandra; Burbank, Malcolm; Paszczynski, Andrzej J
Mutagenesis in Escherichia coli lacking catalase.
Escherichia coli K-12 strains completely lacking catalase activity due to mutations in katG, katE, and katF genes were constructed in order to assess the role of hydrogen peroxide in mutagenesis. Mutagenesis was monitored by selecting forward mutations to L-arabinose resistance. Lethality was measured at experimental conditions equivalent to those of the mutant yield by using a mixed culture of pairs of isogenic strains distinguished by their differential nutritional requirements. Deficiency in katG, katE, and katF genes leads to an enhanced spontaneous mutation rate as well as an enhanced sensitivity to both the lethal and mutagenic effects of hydrogen peroxide or an H2O2-generating mixture of compounds, such as coffee. To compare further the responses of the catalase-deficient bacteria to those of catalase-proficient counterparts, other genotoxins were analyzed. Both catalase-deficient and catalase-proficient strains were equally mutated by MMS, 4-NQO, and ultraviolet light. It is concluded that the bacterial strains and the mutagenicity tests described in the paper represent a useful tool to study the role of H2O2 in mutagenesis. PMID:2192882
Abril, N; Pueyo, C
Protection of Bacillus pumilus Spores by Catalases
Bacillus pumilus SAFR-032, isolated at spacecraft assembly facilities of the National Aeronautics and Space Administration Jet Propulsion Laboratory, is difficult to kill by the sterilization method of choice, which uses liquid or vapor hydrogen peroxide. We identified two manganese catalases, YjqC and BPUM_1305, in spore protein extracts of several B. pumilus strains by using PAGE and mass spectrometric analyses. While the BPUM_1305 catalase was present in six of the B. pumilus strains tested, YjqC was not detected in ATCC 7061 and BG-B79. Furthermore, both catalases were localized in the spore coat layer along with laccase and superoxide dismutase. Although the initial catalase activity in ATCC 7061 spores was higher, it was less stable over time than the SAFR-032 enzyme. We propose that synergistic activity of YjqC and BPUM_1305, along with other coat oxidoreductases, contributes to the enhanced resistance of B. pumilus spores to hydrogen peroxide. We observed that the product of the catalase reaction, gaseous oxygen, forms expanding vesicles on the spore surface, affecting the mechanical integrity of the coat layer, resulting in aggregation of the spores. The accumulation of oxygen gas and aggregations may play a crucial role in limiting further exposure of Bacilli spore surfaces to hydrogen peroxide or other toxic chemicals when water is present.
Checinska, Aleksandra; Burbank, Malcolm
Discovery of Catalases in Members of the Chlamydiales Order
Catalase is an important virulence factor for survival in macrophages and other phagocytic cells. In Chlamydiaceae, no catalase had been described so far. With the sequencing and annotation of the full genomes of Chlamydia-related bacteria, the presence of different catalase-encoding genes has been documented. However, their distribution in the Chlamydiales order and the functionality of these catalases remain unknown. Phylogeny of chlamydial catalases was inferred using MrBayes, maximum likelihood, and maximum parsimony algorithms, allowing the description of three clade 3 and two clade 2 catalases. Only monofunctional catalases were found (no catalase-peroxidase or Mn-catalase). All presented a conserved catalytic domain and tertiary structure. Enzymatic activity of cloned chlamydial catalases was assessed by measuring hydrogen peroxide degradation. The catalases are enzymatically active with different efficiencies. The catalase of Parachlamydia acanthamoebae is the least efficient of all (its catalytic activity was 2 logs lower than that of Pseudomonas aeruginosa). Based on the phylogenetic analysis, we hypothesize that an ancestral class 2 catalase probably was present in the common ancestor of all current Chlamydiales but was retained only in Criblamydia sequanensis and Neochlamydia hartmannellae. The catalases of class 3, present in Estrella lausannensis and Parachlamydia acanthamoebae, probably were acquired by lateral gene transfer from Rhizobiales, whereas for Waddlia chondrophila they likely originated from Legionellales or Actinomycetales. The acquisition of catalases on several occasions in the Chlamydiales suggests the importance of this enzyme for the bacteria in their host environment.
Rusconi, Brigida
Confirmation that catalase is a glycoprotein.
Catalases which had been purified from the livers of mouse, rat and guinea pig were subjected to mild periodate oxidation followed by reduction with sodium boro[3H]hydride in order to test for the presence of sialic acid. A radioactively labelled moiety resulted, which behaved as a derivative of N-acetyl neuraminic acid during mild acid hydrolysis, neuraminidase treatment, ion exchange chromatography and paper chromatography. It is concluded that mammalian catalases are glycoproteins, and possess variable amounts of N-acetyl neuraminic acid in their carbohydrate moiety. PMID:3017350
Pegg, M; Crane, D; Masters, C
21 CFR 173.135 - Catalase derived from Micrococcus lysodeikticus.
Code of Federal Regulations, 2013 CFR
...CONTINUED) FOOD FOR HUMAN CONSUMPTION (CONTINUED) SECONDARY DIRECT FOOD ADDITIVES PERMITTED IN FOOD FOR HUMAN CONSUMPTION Enzyme Preparations and Microorganisms § 173.135 Catalase derived from Micrococcus lysodeikticus. Bacterial catalase...
21 CFR 184.1034 - Catalase (bovine liver).
Code of Federal Regulations, 2010 CFR
...2010-01-01 2009-04-01 true Catalase (bovine liver). 184.1034 Section 184.1034 Food and...Affirmed as GRAS § 184.1034 Catalase (bovine liver). (a) Catalase (bovine liver) (CAS Reg. No. 81457-95-6) is an...
21 CFR 184.1034 - Catalase (bovine liver).
Code of Federal Regulations, 2010 CFR
...2009-04-01 2009-04-01 false Catalase (bovine liver). 184.1034 Section 184.1034 Food and...Affirmed as GRAS § 184.1034 Catalase (bovine liver). (a) Catalase (bovine liver) (CAS Reg. No. 81457-95-6) is an...
A Eukaryote without Catalase-Containing Microbodies: Neurospora crassa Exhibits a Unique Cellular Distribution of Its Four Catalases
Microbodies usually house catalase to decompose hydrogen peroxide generated within the organelle by the action of various oxidases. Here we have analyzed whether peroxisomes (i.e., catalase-containing microbodies) exist in Neurospora crassa. Three distinct catalase isoforms were identified by native catalase activity gels under various peroxisome-inducing conditions. Subcellular fractionation by density gradient centrifugation revealed that most of the spectrophotometrically measured activity
Wolfgang Schliebs; Christian W�rtz; Wolf-Hubert Kunau; Marten Veenhuis; Hanspeter Rottensteiner
Ferrous complexes in the catalase reaction
R�sum� L'auteur critique l'id�e que la polarographie d�montra l'existence de complexes d'h�matine ferreuse avec le peroxide, actifs dans la catalyse. D'autres exp�riences montrent que les d�riv�s de l'h�matine et du peroxyde sont instables et provoquent la d�gradation de la porphyrine. Les complexes de la catalase et du peroxyde ne sont pas actifs polarographiquement et contiennent probablement le fer � l'�tat
P. Nicholls
Catalytic scavenging of peroxynitrite by catalase
Peroxynitrite (ONOO?\\/ONOOH), the product of the diffusion controlled reaction between nitric oxide (NO) and superoxide anion (O2-), is a strong oxidizing and nitrating agent. Several heme proteins react rapidly with peroxynitrite, some of them catalyze its decomposition. In this work we found, contrary to previous reports, that catalase, a ferriheme enzyme, catalytically scavenges peroxynitrite. The second-order reaction rate constants of
Lidia Gebicka; Joanna Didik
Probing the binding of flavonoids to catalase by molecular spectroscopy
NASA Astrophysics Data System (ADS)
The binding of flavonoids (quercetin and myricetin) to catalase was investigated by fluorescence and circular dichroism (CD) techniques under physiological conditions. The binding parameters and binding mode between flavonoids and catalase were determined, and the results of synchronous fluorescence spectra and CD indicated a conformational change of catalase with addition of flavonoids. The effect of both Cu 2+ and vitamin C on the binding constant of flavonoid-catalase was also examined. The experiment data show that the difference of the structure characteristics of quercetin and myricetin has a significant effect on their binding affinity for catalase.
Zhu, Jingfeng; Zhang, Xia; Li, Daojin; Jin, Jing
The Stringent Response Controls Catalases in Pseudomonas aeruginosa and Is Required for Hydrogen Peroxide and Antibiotic Tolerance
Pseudomonas aeruginosa, a human opportunistic pathogen, possesses a number of antioxidant defense enzymes under the control of multiple regulatory systems. We recently reported that inactivation of the P. aeruginosa stringent response (SR), a starvation stress response controlled by the alarmone (p)ppGpp, caused impaired antioxidant defenses and antibiotic tolerance. Since catalases are key antioxidant enzymes in P. aeruginosa, we compared the levels of H2O2 susceptibility and catalase activity in P. aeruginosa wild-type and ?relA ?spoT (?SR) mutant cells. We found that the SR was required for optimal catalase activity and mediated H2O2 tolerance during both planktonic and biofilm growth. Upon amino acid starvation, induction of the SR upregulated catalase activity. Full expression of katA and katB also required the SR, and this regulation occurred through both RpoS-independent and RpoS-dependent mechanisms. Furthermore, overexpression of katA was sufficient to restore H2O2 tolerance and to partially rescue the antibiotic tolerance of ?SR cells. All together, these results suggest that the SR regulates catalases and that this is an important mechanism in protecting nutrient-starved and biofilm bacteria from H2O2- and antibiotic-mediated killing.
Khakimova, Malika; Ahlgren, Heather G.; Harrison, Joe J.; English, Ann M.
The stringent response controls catalases in Pseudomonas aeruginosa and is required for hydrogen peroxide and antibiotic tolerance.
Pseudomonas aeruginosa, a human opportunistic pathogen, possesses a number of antioxidant defense enzymes under the control of multiple regulatory systems. We recently reported that inactivation of the P. aeruginosa stringent response (SR), a starvation stress response controlled by the alarmone (p)ppGpp, caused impaired antioxidant defenses and antibiotic tolerance. Since catalases are key antioxidant enzymes in P. aeruginosa, we compared the levels of H2O2 susceptibility and catalase activity in P. aeruginosa wild-type and ?relA ?spoT (?SR) mutant cells. We found that the SR was required for optimal catalase activity and mediated H2O2 tolerance during both planktonic and biofilm growth. Upon amino acid starvation, induction of the SR upregulated catalase activity. Full expression of katA and katB also required the SR, and this regulation occurred through both RpoS-independent and RpoS-dependent mechanisms. Furthermore, overexpression of katA was sufficient to restore H2O2 tolerance and to partially rescue the antibiotic tolerance of ?SR cells. All together, these results suggest that the SR regulates catalases and that this is an important mechanism in protecting nutrient-starved and biofilm bacteria from H2O2- and antibiotic-mediated killing. PMID:23457248
Khakimova, Malika; Ahlgren, Heather G; Harrison, Joe J; English, Ann M; Nguyen, Dao
Molecular Characterization of a Catalase from Hydra vulgaris
Catalase, an antioxidant and hydroperoxidase enzyme protects the cellular environment from harmful effects of hydrogen peroxide by facilitating its degradation to oxygen and water. Molecular information on a cnidarian catalase and/or peroxidase is, however, limited. In this work an apparent full length cDNA sequence coding for a catalase (HvCatalase) was isolated from Hydra vulgaris using 3�- and 5�- (RLM) RACE approaches. The 1859 bp HvCatalase cDNA included an open reading frame of 1518 bp encoding a putative protein of 505 amino acids with a predicted molecular mass of 57.44 kDa. The deduced amino acid sequence of HvCatalase contained several highly conserved motifs including the heme-ligand signature sequence RLFSYGDTH and the active site signature FXRERIPERVVHAKGXGA. A comparative analysis showed the presence of conserved catalytic amino acids [His(71), Asn(145), and Tyr(354)] in HvCatalase as well. Homology modeling indicated the presence of the conserved features of mammalian catalase fold. Hydrae exposed to thermal, starvation, metal and oxidative stress responded by regulating its catalase mRNA transcription. These results indicated that the HvCatalase gene is involved in the cellular stress response and (anti)oxidative processes triggered by stressor and contaminant exposure.
Dash, Bhagirathi; Phillips, Timothy D.
Relation of Catalase to Substrate Utilization by Mycoplasma pneumoniae
No catalase activity was detected in four strains of glucose-grown Mycoplasma pneumoniae at any time during the replication of the organism. Exogenous catalase dramatically increased the O2 uptake with glycerol, presumably by releasing inhibition caused by hydrogen peroxide. The effect of added catalase on the O2 uptake of washed organisms with glucose as substrate was moderate and variable in degree. The production of hydrogen peroxide was demonstrated by the quantitative enzymatic assay for inorganic peroxide and by the fact that added pyruvate, which is non-enzymatically oxidized by H2O2 to acetic acid and CO2 could mimic the action of catalase.
Low, I. E.; Eaton, M. D.; Proctor, P.
A Eukaryote without Catalase-Containing Microbodies: Neurospora crassa Exhibits a Unique Cellular Distribution of Its Four Catalases�
Microbodies usually house catalase to decompose hydrogen peroxide generated within the organelle by the action of various oxidases. Here we have analyzed whether peroxisomes (i.e., catalase-containing microbodies) exist in Neurospora crassa. Three distinct catalase isoforms were identified by native catalase activity gels under various peroxisome-inducing conditions. Subcellular fractionation by density gradient centrifugation revealed that most of the spectrophotometrically measured activity was present in the light upper fractions, with an additional small peak coinciding with the peak fractions of HEX-1, the marker protein for Woronin bodies, a compartment related to the microbody family. However, neither in-gel assays nor monospecific antibodies generated against the three purified catalases detected the enzymes in any dense organellar fraction. Furthermore, staining of an N. crassa wild-type strain with 3,3?-diaminobenzidine and H2O2 did not lead to catalase-dependent reaction products within microbodies. Nonetheless, N. crassa does possess a gene (cat-4) whose product is most similar to the peroxisomal type of monofunctional catalases. This novel protein indeed exhibited catalase activity, but was not localized to microbodies either. We conclude that N. crassa lacks catalase-containing peroxisomes, a characteristic that is probably restricted to a few filamentous fungi that produce little hydrogen peroxide within microbodies.
Schliebs, Wolfgang; Wurtz, Christian; Kunau, Wolf-Hubert; Veenhuis, Marten; Rottensteiner, Hanspeter
Catalases Are NAD(P)H-Dependent Tellurite Reductases
Reactive oxygen species damage intracellular targets and are implicated in cancer, genetic disease, mutagenesis, and aging. Catalases are among the key enzymatic defenses against one of the most physiologically abundant reactive oxygen species, hydrogen peroxide. The well-studied, heme-dependent catalases accelerate the rate of the dismutation of peroxide to molecular oxygen and water with near kinetic perfection. Many catalases also bind the cofactors NADPH and NADH tenaciously, but, surprisingly, NAD(P)H is not required for their dismutase activity. Although NAD(P)H protects bovine catalase against oxidative damage by its peroxide substrate, the catalytic role of the nicotinamide cofactor in the function of this enzyme has remained a biochemical mystery to date. Anions formed by heavy metal oxides are among the most highly reactive, natural oxidizing agents. Here, we show that a natural isolate of Staphylococcus epidermidis resistant to tellurite detoxifies this anion thanks to a novel activity of its catalase, and that a subset of both bacterial and mammalian catalases carry out the NAD(P)H-dependent reduction of soluble tellurite ion (TeO32?) to the less toxic, insoluble metal, tellurium (Te�), in vitro. An Escherichia coli mutant defective in the KatG catalase/peroxidase is sensitive to tellurite, and expression of the S. epidermidis catalase gene in a heterologous E. coli host confers increased resistance to tellurite as well as to hydrogen peroxide in vivo, arguing that S. epidermidis catalase provides a physiological line of defense against both of these strong oxidizing agents. Kinetic studies reveal that bovine catalase reduces tellurite with a low Michaelis-Menten constant, a result suggesting that tellurite is among the natural substrates of this enzyme. The reduction of tellurite by bovine catalase occurs at the expense of producing the highly reactive superoxide radical.
Calderon, Ivan L.; Arenas, Felipe A.; Perez, Jose Manuel; Fuentes, Derie E.; Araya, Manuel A.; Saavedra, Claudia P.; Tantalean, Juan C.; Pichuantes, Sergio E.; Youderian, Philip A.; Vasquez, Claudio C.
Genes Important for Catalase Activity in Enterococcus faecalis
Little in general is known about how heme proteins are assembled from their constituents in cells. The Gram-positive bacterium Enterococcus faecalis cannot synthesize heme and does not depend on it for growth. However, when supplied with heme in the growth medium the cells can synthesize two heme proteins; catalase (KatA) and cytochrome bd (CydAB). To identify novel factors important for catalase biogenesis libraries of E. faecalis gene insertion mutants were generated using two different types of transposons. The libraries of mutants were screened for clones deficient in catalase activity using a colony zymogram staining procedure. Analysis of obtained clones identified, in addition to katA (encoding the catalase enzyme protein), nine genes distributed over five different chromosomal loci. No factors with a dedicated essential role in catalase biogenesis or heme trafficking were revealed, but the results indicate the RNA degradosome (srmB, rnjA), an ABC-type oligopeptide transporter (oppBC), a two-component signal transducer (etaR), and NADH peroxidase (npr) as being important for expression of catalase activity in E. faecalis. It is demonstrated that catalase biogenesis in E. faecalis is independent of the CydABCD proteins and that a conserved proline residue in the N-terminal region of KatA is important for catalase assembly.
Baureder, Michael; Hederstedt, Lars
Microcalorimetric studies on catalase reaction and inhibition of catalase by cyanide ion
As Chance et al. [1�3] proposed, the decomposition of hydrogen peroxide catalyzed by catalase is an overall first-order reaction. In this paper, we have studied this enzyme-catalyzed reaction with a thermokinetic method. The rate constant and the molar reaction enthalpy of this reaction have been measured. At 310.15K and pH=8.2, kcat=1.75�106lmol?1s?1, ?rHm=88.99kJmol?1. Furthermore, we have studied the competitive inhibition of
Wang Zhiyong; Wang Cunxin; Qu Songsheng
Identification of Two Catalases in Azotobacter vinelandii: a KatG Homologue and a Novel Bacterial Cytochrome c Catalase, CCCAv? �
Azotobacter vinelandii produces two detectable catalases during growth on minimal medium. The heat-labile catalase expressed during exponential growth phase was identified as a KatG homologue by liquid chromatography-tandem mass spectrometry (LC-MS/MS) using a mixed protein sample. The second catalase was heat resistant and had substantial residual activity after treatment at 90�C. This enzyme was purified by anion-exchange and size exclusion chromatography and was found to exhibit strong absorption at 407 nm, which is often indicative of associated heme moieties. The purified protein was fragmented by proteinase K and identified by LC-MS/MS. Some identity was shared with the MauG/bacterial cytochrome c peroxidase (BCCP) protein family, but the enzyme exhibited a strong catalase activity never before observed in this family. Because two putative c-type heme sites (CXXCH) were predicted in the peptide sequence and were demonstrated experimentally, the enzyme was designated a cytochrome c catalase (CCCAv). However, the local organization of the CCCAv heme motifs differed significantly from that of the BCCPs as the sites were confined to the C-terminal half of the catalase. A possible Ca2+ binding motif, previously described in the BCCPs, is also present in the CCCAv peptide sequence. Some instability in the presence of EGTA was observed. Expression of the catalase was abolished in cccA mutants, resulting in a nearly 8,700-fold reduction in peroxide resistance in stationary phase.
Sandercock, James R.; Page, William J.
The catalase activity of diiron adenine deaminase
Adenine deaminase (ADE) from the amidohydrolase superfamily (AHS) of enzymes catalyzes the conversion of adenine to hypoxanthine and ammonia. Enzyme isolated from Escherichia coli was largely inactive toward the deamination of adenine. Molecular weight determinations by mass spectrometry provided evidence that multiple histidine and methionine residues were oxygenated. When iron was sequestered with a metal chelator and the growth medium supplemented with Mn2+ before induction, the post-translational modifications disappeared. Enzyme expressed and purified under these conditions was substantially more active for adenine deamination. Apo-enzyme was prepared and reconstituted with two equivalents of FeSO4. Inductively coupled plasma mass spectrometry and M�ssbauer spectroscopy demonstrated that this protein contained two high-spin ferrous ions per monomer of ADE. In addition to the adenine deaminase activity, [FeII/FeII]-ADE catalyzed the conversion of H2O2 to O2 and H2O. The values of kcat and kcat/Km for the catalase activity are 200 s?1 and 2.4 � 104 M?1 s?1, respectively. [FeII/FeII]-ADE underwent more than 100 turnovers with H2O2 before the enzyme was inactivated due to oxygenation of histidine residues critical for metal binding. The iron in the inactive enzyme was high-spin ferric with gave = 4.3 EPR signal and no evidence of anti-ferromagnetic spin-coupling. A model is proposed for the disproportionation of H2O2 by [FeII/FeII]-ADE that involves the cycling of the binuclear metal center between the di-ferric and di-ferrous oxidation states. Oxygenation of active site residues occurs via release of hydroxyl radicals. These findings represent the first report of redox reaction catalysis by any member of the AHS.
Kamat, Siddhesh S; Holmes-Hampton, Gregory P; Bagaria, Ashima; Kumaran, Desigan; Tichy, Shane E; Gheyi, Tarun; Zheng, Xiaojing; Bain, Kevin; Groshong, Chris; Emtage, Spencer; Sauder, J Michael; Burley, Stephen K; Swaminathan, Subramanyam; Lindahl, Paul A; Raushel, Frank M
Pulse radiolysis of catalase in solution�II. Reactions of primary products from water radiolysis with catalase
NASA Astrophysics Data System (ADS)
The mechanism of the reaction of catalase with e -aq, H and OH has been studied by pulse radiolysis at room temperature. Some evidences have been found that e -eq/H react with porphyrin ring of catalase to form ?-radical without reduction of heme iron within investigated time span of 1 s after the pulse. OH radicals react mainly with the protein moiety of the enzyme but the formation and decay of compound I, an intermediate of the catalytic reaction of catalase can be observed as well.
G?bicka, Lidia; G?bicki, Jerzy L.
A Laboratory Experiment of the Purification of Catalase.
ERIC Educational Resources Information Center
Describes a simple method for purifying catalase for the study of proteins. Procedures are systematically and diagramatically presented. Also identifies polyacrylamide gel electrophoresis, kinetic studies, and apparent molecular weight determination as possible techniques to be used in studying proteins. (ML)
Busquets, Montserrat; Franco, Rafael
Comparative study of catalase-peroxidases (KatGs)
Catalase-peroxidases or KatGs from seven different organisms, including Archaeoglobus fulgidus,Bacillus stearothermophilus, Burkholderia pseudomallei, Escherichia coli, Mycobacterium tuberculosis, Rhodobacter capsulatus and Synechocystis PCC 6803, have been characterized to provide a comparative picture of their respective properties. Collectively, the enzymes exhibit similar turnover rates with the catalase and peroxidase reactions varying between 4900 and 15,900s?1 and 8�25s?1, respectively. The seven enzymes also
Rahul Singh; Ben Wiseman; Taweewat Deemagarn; Vikash Jha; Jacek Switala; Peter C. Loewen
The stability of catalase in the reaction of hydrogen peroxide decomposition was studied in the presence of some potential stabilizers: ethylene glycol, glycerol, fructose, sucrose, fucose, ribitol and 2 nitro phenyl- #-D-galacto-pyranosid (niphegal). The sucrose, ethylene glycol and glycerol showed the best performance for long-term storage at 30�C. The stability of catalase increases with the amount of additive used.
Madalina Nita; Adina Raducan; Mihaela Puiu; D. Oancea
Comparative immunological study of catalases in the genus Micrococcus
Double immunodiffusion tests were performed with crude extracts from various Micrococcus species and antisera against catalase of Micrococcus luteus CCM 169. Cell-free extracts of M. lylae ATCC 27566 exhibited good cross-reaction. Cell-free extracts or catalase enriched preparations of M. varians reacted very weakly and no reaction has been found with preparations of M. kristinae, M. nishinomiyaensis, M. roseus and M.
Manfred Rupprecht; Karl-Heinz Schleifer
Characterization of a cDNA encoding cottonseed catalase.
A 1.7 kb cDNA clone was isolated from our lambda gt11 library constructed from poly(A) RNA of 24-h-old cotyledons. The cDNA encodes a full-length catalase peptide (492 amino acid residues). The calculated molecular mass is 56,800, similar to that determined for purified enzyme (57,000 SDS-PAGE). Among higher plant catalases, this cotton catalase shows the highest amino acid sequence identity (85%) to the subunit of homotetrameric maize CAT 1, a developmental counterpart to the homotetrameric CAT A isoform of cotton seeds. Comparison of sequences from cotton, sweet potato, maize CAT 1, and yeast with bovine catalase revealed that the amino acid residues and regions that are involved in catalytic activity and/or required to maintain basic catalase structure, are highly conserved. The C-terminus region, which has the lowest nucleotide sequence identity between plant and mammalian catalases, does not terminate with a tripeptide, S-K/R/H-L, a putative targeting signal for peroxisomal proteins. PMID:2364113
Ni, W; Turley, R B; Trelease, R N
Formulation and characterization of catalase in albumin microspheres.
Catalase in albumin microspheres were formulated for intravenous administration to antagonize the effects of over-production of reactive oxygenated species (ROS) such as hydrogen peroxide (H(2)O(2)) in septic shock. The aim was to increase effective half-life of catalase and take advantage of the phagocytic uptake of the encapsulated catalase by the vascular endothelium. Catalase microspheres were prepared by spray-drying. The microspheres were evaluated for particle size, particle shape and surface morphology by scanning electron microscopy (SEM), drug encapsulation efficiency, chemical stability, thermal stability and in vitro drug release characteristics. The microspheres had a mean particle size of 4.7 +/- 2 microm, optimal for phagocytic uptake, as demonstrated by Makino et al. SEM revealed that microspheres were spherical with smooth surface morphology. An encapsulation efficiency of 91.5 +/- 3% was achieved and the encapsulated catalase was chemically and thermally stable. Application of in vitro drug release data to the Higuchi kinetic equation indicated matrix diffusion-controlled catalase release from albumin microspheres. PMID:18821261
Siwale, Rodney C; Oettinger, Carl W; Pai, S Balakrishna; Addo, Richard; Uddin, Nasir; Siddig, Aladin; D'Souza, Martin J
Pseudomonas syringae Catalases Are Collectively Required for Plant Pathogenesis
The bacterial pathogen Pseudomonas syringae pv. tomato DC3000 must detoxify plant-produced hydrogen peroxide (H2O2) in order to survive in its host plant. Candidate enzymes for this detoxification include the monofunctional catalases KatB and KatE and the bifunctional catalase-peroxidase KatG of DC3000. This study shows that KatG is the major housekeeping catalase of DC3000 and provides protection against menadione-generated endogenous H2O2. In contrast, KatB rapidly and substantially accumulates in response to exogenous H2O2. Furthermore, KatB and KatG have nonredundant roles in detoxifying exogenous H2O2 and are required for full virulence of DC3000 in Arabidopsis thaliana. Therefore, the nonredundant ability of KatB and KatG to detoxify plant-produced H2O2 is essential for the bacteria to survive in plants. Indeed, a DC3000 catalase triple mutant is severely compromised in its ability to grow in planta, and its growth can be partially rescued by the expression of katB, katE, or katG. Interestingly, our data demonstrate that although KatB and KatG are the major catalases involved in the virulence of DC3000, KatE can also provide some protection in planta. Thus, our results indicate that these catalases are virulence factors for DC3000 and are collectively required for pathogenesis.
Guo, Ming; Block, Anna; Bryan, Crystal D.; Becker, Donald F.
The catalase activity of diiron adenine deaminase
Adenine deaminase (ADE) from the amidohydrolase superfamily (AHS) of enzymes catalyzes the conversion of adenine to hypoxanthine and ammonia. Enzyme isolated from Escherichia coli was largely inactive toward the deamination of adenine. Molecular weight determinations by mass spectrometry provided evidence that multiple histidine and methionine residues were oxygenated. When iron was sequestered with a metal chelator and the growth medium supplemented with Mn{sup 2+} before induction, the post-translational modifications disappeared. Enzyme expressed and purified under these conditions was substantially more active for adenine deamination. Apo-enzyme was prepared and reconstituted with two equivalents of FeSO{sub 4}. Inductively coupled plasma mass spectrometry and Moessbauer spectroscopy demonstrated that this protein contained two high-spin ferrous ions per monomer of ADE. In addition to the adenine deaminase activity, [Fe{sup II}/Fe{sup II}]-ADE catalyzed the conversion of H{sub 2}O{sub 2} to O{sub 2} and H{sub 2}O. The values of k{sub cat} and k{sub cat}/K{sub m} for the catalase activity are 200 s{sup -1} and 2.4 x 10{sup 4} M{sup -1} s{sup -1}, respectively. [Fe{sup II}/Fe{sup II}]-ADE underwent more than 100 turnovers with H{sub 2}O{sub 2} before the enzyme was inactivated due to oxygenation of histidine residues critical for metal binding. The iron in the inactive enzyme was high-spin ferric with g{sub ave} = 4.3 EPR signal and no evidence of anti-ferromagnetic spin-coupling. A model is proposed for the disproportionation of H{sub 2}O{sub 2} by [Fe{sup II}/Fe{sup II}]-ADE that involves the cycling of the binuclear metal center between the di-ferric and di-ferrous oxidation states. Oxygenation of active site residues occurs via release of hydroxyl radicals. These findings represent the first report of redox reaction catalysis by any member of the AHS.
Kamat S. S.; Swaminathan S.; Holmes-Hampton, G. P.; Bagaria, A.; Kumaran, D.; Tichy, S. E.; Gheyi, T.; Zheng, X.; Bain, K.; Groshong, C.; Emtage, S.; Sauder, J. M.; Burley, S. K.; Lindahl, P. A.; Raushel, F. M.
Inhibition of Activity of Catalase from Potato Tubers by Salicylic and Succinic Acids
It was found that salicylic acid inhibits the activity of catalase from potato tubers in vitro. Succinic acid suppressed catalase activity at the same concentrations that salicylate; however, its effect was more long-term. Bovine catalase was less sensitive to salicylic and succinic acids than potato catalase. In the past years, the attention of researchers has been attracted to studying the
Ya. S. Panina; N. I. Vasyukova; O. L. Ozeretskovskaya
Characterization of catalase transcripts and their differential expression in maize.
In maize, the three unlinked catalase (EC 184.108.40.206) structural genes (Cat1, Cat2 and Cat3) are differentially expressed temporally, spatially and in response to environmental signals in the developing seedling. In order to understand more fully the molecular mechanisms involved in catalase gene expression, full-length cDNA clones representing the maize Cat1, Cat2 and Cat3 transcripts were isolated and characterized. DNA sequence analysis confirmed that each cDNA encodes a unique catalase protein. Gene-specific probes for the three maize catalase cDNAs were isolated and used to probe blots of poly(A)+ RNA isolated from various maize tissues. Cat1 mRNA was found in scutella, milky endosperm of immature kernels, leaves and epicotyls. The Cat2 mRNA was present primarily in post-germinative scutella, with lower levels in leaves and epicotyls. Cat3 mRNA was detected primarily in epicotyls and, to a lesser extent, in leaves and scutella. The gene-specific probes hybridized with maize genomic DNA blots in simple, but unique patterns, indicating that there is one, or a very few copies of each catalase gene. The coding region of the Cat3 cDNA comprised 66% G + C, which led to a strong codon usage bias in this gene. This codon bias was also seen with the Cat2 transcripts, but not with those for Cat1. A high degree of similarity was found between the maize catalase nucleic acid and deduced amino-acid sequences and those of sweet potato and rat liver catalase. PMID:2461221
Redinbaugh, M G; Wadsworth, G J; Scandalios, J G
Increased microglial catalase activity in multiple sclerosis grey matter.
Chronic demyelination, on-going inflammation, axonal loss and grey matter neuronal injury are likely pathological processes that contribute to disease progression in multiple sclerosis (MS). Although the precise contribution of each process and their aetiological substrates is not fully known, recent evidence has implicated oxidative damage as a major cause of tissue injury in MS. The degree of tissue injury caused by oxidative molecules, such as reactive oxygen species (ROS), is balanced by endogenous anti-oxidant enzymes which detoxify ROS. Understanding endogenous mechanisms which protect the brain against oxidative injury in MS is important, since enhancing anti-oxidant responses is a major therapeutic strategy for preventing irreversible tissue injury in the disease. Our aims were to determine expression and activity levels of the hydrogen peroxide-reducing enzyme catalase in MS grey matter (GM). In MS GM, a catalase enzyme activity was elevated compared to control GM. We measured catalase protein expression by immune dot-blotting and catalase mRNA by a real-time polymerase chain reaction (RT-PCR). Protein analysis studies showed a strong positive correlation between catalase and microglial marker IBA-1 in MS GM. In addition, calibration of catalase mRNA level with reference to the microglial-specific transcript AIF-1 revealed an increase in this transcript in MS. This was reflected by the extent of HLA-DR immunolabeling in MS GM which was significantly elevated compared to control GM. Collectively, these observations provide evidence that microglial catalase activity is elevated in MS grey matter and may be an important endogenous anti-oxidant defence mechanism in MS. PMID:24602691
Gray, Elizabeth; Kemp, Kevin; Hares, Kelly; Redondo, Julianna; Rice, Claire; Scolding, Neil; Wilkins, Alastair
Catalase-negative Staphylococcus lugdunensis strain with a novel point mutation in the catalase gene isolated from a patient with chronic suppurative otitis media.
This report describes the results of the sequence analysis of a methicillin-susceptible strain of catalase-negative Staphylococcus lugdunensis. Molecular characterization of the deduced sequence revealed a novel point mutation in the catalase gene. To our knowledge, this is the first report of a catalase-negative S. lugdunensis strain, although catalase-negative isolates of Staphylococcus aureus and Staphylococcus epidermidis have been previously reported. PMID:23345293
Lu, Yong; Wang, Yiping; Ling, Buzhi; Ke, Xianfu; Ying, Jianfei; Yu, Yanhong; He, Mingyang; Li, Xiangyang
A rapid method for detection of catalase-positive and catalase-negative bacteria based on monitoring of hydrogen peroxide evolution at a composite peroxidase biosensor
The rapid detection of catalase-positive and catalase-negative bacteria in complex culture media has been accomplished by monitoring of hydrogen peroxide consumption or generation with a graphite�Teflon�peroxidase�ferrocene composite electrode. Escherichia coli and Streptococcus pneumoniae have been used as model catalase-positive and catalase-negative bacteria, respectively. Hydrogen peroxide evolution was amperometrically measured at 0.00V. Experimental conditions, including the working solution composition, the incubation
B. Serra; J. Zhang; M. D. Morales; A. Guzm�n-V�zquez de Prada; A. J. Reviejo; J. M. Pingarr�n
Superoxide dismutase and catalase activities in Photobacterium damselae ssp. piscicida.
The ability of a set of Photobacterium damselae ssp. piscicida strains isolated from different fish species to produce different superoxide dismutase (SOD) and catalase enzymes was determined. Unlike other bacterial pathogens, P. damselae ssp. piscicida is not able to produce different isoforms of SOD or catalase containing different metal cofactors when cultured under oxidative stress induced by hydrogen peroxide or methyl viologen, or under iron depleted conditions. However, iron content of the growth medium influenced the levels of SOD and catalase activity in cells, these levels decreasing with iron availability of the medium. Comparison of virulent and non-virulent strains of P. damselae ssp. piscicida showed similar contents of SOD, but higher levels of catalase were detected in cells of the virulent strain. Incubation of bacteria with sole, Solea senegalensis (Kaup), phagocytes has shown that survival rates range from 19% to 62%, these rates being higher for the virulent strain. The increased levels of catalase activity detected in the virulent strain indicates a possible role for this enzyme in bacterial survival. PMID:16768716
D�az-Rosales, P; Chabrill�n, M; Arijo, S; Martinez-Manzanares, E; Mori�igo, M A; Balebona, M C
[Influence of magnetic field on hydrogen peroxide decomposition by catalase].
The rate constants of H2O2 decomposition equal to 0.45 X 10(7) M-1S-1 per hem and Michaelis constants higher than 0.3 M were found by gas volume meter and spectrophotometer methods for purified preparations of catalase from bovine liver. Unlike the results obtained earlier the magnetic field with induction 0.65 T and 0.012 T does not affect the constant rate within 3%. It was found with the substrate concentration less than 0.01 M when the classical catalase mechanism was observed and with higher concentration of the substrate up to 0.7 M when the catalase inhibition by H2O2 played an important role. PMID:6830896
Vanag, V K; Kuznetsov, A N; Piruzian, L A
Kinetics of hydrogen peroxide decomposition by catalase: hydroxylic solvent effects.
The effect of water-alcohol (methanol, ethanol, propan-1-ol, propan-2-ol, ethane-1,2-diol and propane-1,2,3-triol) binary mixtures on the kinetics of hydrogen peroxide decomposition in the presence of bovine liver catalase is investigated. In all solvents, the activity of catalase is smaller than in water. The results are discussed on the basis of a simple kinetic model. The kinetic constants for product formation through enzyme-substrate complex decomposition and for inactivation of catalase are estimated. The organic solvents are characterized by several physical properties: dielectric constant (D), hydrophobicity (log P), concentration of hydroxyl groups ([OH]), polarizability (?), Kamlet-Taft parameter (?) and Kosower parameter (Z). The relationships between the initial rate, kinetic constants and medium properties are analyzed by linear and multiple linear regression. PMID:22565543
Raducan, Adina; Cantemir, Anca Ruxandra; Puiu, Mihaela; Oancea, Dumitru
Negative regulation of catalase gene expression in hepatoma cells.
For an understanding of the molecular basis of the marked decrease in catalase activity of various tumor cells, expression of the catalase gene was studied in rat and human hepatoma cell lines and in rat liver, which was used as a control with high activity. RNA blot hybridization profiles and run-on assays indicated that the decrease in catalase activity was due to depression of catalase gene transcription. Chloramphenicol acetyltransferase (CAT) assays for the fragments with various lengths of the 5'-flanking region (up to -4.5 kb from the ATG codon) of the catalase gene revealed the presence of several cis-acting elements involved in the negative regulation of transcription. The most-upstream element with the strongest activity (-3504 to -3364 bp), when linked to the catalase promoter region (-126 bp) of the CAT construct and subjected to an in vitro transcription assay, did not yield transcripts in experiments with the hepatoma nuclear extract, whereas the unlinked template did yield transcripts. A gel shift competition assay using hepatoma nuclear extract showed the core sequence of the silencer element to be 5'-TGGGGGGAG-3'. A homology search found that the same core sequence was also present in 5'-flanking regions of the albumin gene and of some other liver enzyme genes, the expression of which has been reported to be down regulated in some hepatoma cells. Southwestern (DNA-protein) analysis demonstrated that an approximately 35-kDa nuclear protein bound to the silencer element was present in hepatoma cells but not in rat liver cells. Images
Sato, K; Ito, K; Kohara, H; Yamaguchi, Y; Adachi, K; Endo, H
Biological Response of Lactic Streptococci and Lactobacilli to Catalase1
Addition of catalase to milk cultures of lactic streptococci resulted in increased rates of acid production, although it had no effect on cultures of lactobacilli. Milk cultures of both streptococci and lactobacilli produced detectable amounts of peroxide, which reached a maximum level in the early period of acid production followed by a drastic decrease as the acid production increased. Pyruvate and reduced glutathione decreased the amount of peroxide formed, but had little effect on acid production by the streptococci. Ferrous sulfate prevented the accumulation of peroxide and stimulated the rate of acid production by the streptococci to a greater extent than did catalase.
Gilliland, S. E.; Speck, M. L.
Catalase-negative Escherichia coli isolated from blood.
A catalase-negative variant of Escherichia coli was isolated from the blood of a patient with acute leukemia who had been treated with various antibiotics and gentamicin. This small-colony variant grew almost as actively under anaerobic conditions as its large-colony revertant or E. coli NIHJ JC-2. The variant was resistant to gentamicin, in contrast with the revertant. Streptomycin and hemin stimulated growth of the variant slightly. With repeated subculturing the variant tended to increase slightly in colony size with coincident recovery of weak catalase production. The possibility that such a variant may have been induced by gentamicin was indicated. Images
Funada, H; Hattori, K I; Kosakai, N
Improving catalase-based propelled motor endurance by enzyme encapsulation.
Biocatalytic propulsion is expected to play an important role in the future of micromotors as it might drastically increase the number of available fuelling reactions. However, most of the enzyme-propelled micromotors so far reported still rely on the degradation of peroxide by catalase, in spite of being vulnerable to relatively high peroxide concentrations. To overcome this limitation, herein we present a strategy to encapsulate the catalase and to graft the resulting enzyme capsules on motor particles. Significant improvement of the stability in the presence of peroxide and other aggressive agents has been observed. PMID:24964766
Simmchen, Juliane; Baeza, Alejandro; Ruiz-Molina, Daniel; Vallet-Reg�, Maria
Development of lyophilization cycle and effect of excipients on the stability of catalase during lyophilization
Introduction: The purpose of the present study was to screen excipients such as amino acids and non-aqueous solvents for their stabilizing effect on catalase, a model protein, for lyophilization. The present study also includes optimization of lyophilization cycle for catalase formulations, which is essential from the commercial point of view, since lyophilization is an extremely costly process. Materials and Methods: Activity of catalase was determined using catalase activity assay. Differential scanning calorimetry was used to determine eutectic melting temperature of the frozen catalase solution, which is essential for the optimization of lyophilization cycle. Results: When catalase was lyophilized without excipients, it was found that about 65-78% of the initial activity of catalase was lost during the lyophilization process in a concentration dependent manner. The maximum stability of catalase during lyophilization was observed at pH 7.0. Amino acids like alanine, glycine, lysine, serine and 4-hydroxy proline showed strong stabilizing effect on catalase during lyophilization by protecting catalase activity above 95%, whereas valine and cysteine hydrochloride showed destabilizing effect on catalase. Non-aqueous solvents such as dimethyl formamide, dimethyl sulphoxide, polyethylene glycol (PEG) 200, PEG 400, PEG 600 and ethylene glycol also showed destabilizing effect on catalase during lyophilization. Conclusions: In order to prevent loss of catalase activity during lyophilization of catalase, use of amino acids like alanine, glycine, lysine, serine and 4-hydroxy proline in optimum concentration is highly advisable.
Lale, Shantanu V; Goyal, Monu; Bansal, Arvind K
Catalase immobilization in cellulose acetate beads and determination of its hydrogen peroxide decomposition level by using a catalase biosensor.
Catalase enzyme (EC 188.8.131.52) was immobilized by entrapping in cellulose acetate beads. This organic matrix is highly resistant to mechanical stability and can be used under various conditions. Initial studies were conducted to examine the immobilization ability of catalase on the matrix previously activated with a series of reagent normally and the best results were obtained with the beads activated with Ce(SO4)2. In the optimization studies of the immobilized enzyme optimum pH and temperature were found as pH:7.0 (Tris-HCl, 50 mM) and 35 degrees C. In the characterization studies of the immobilized enzyme some parameters such as storage and thermal stability were investigated. Finally, the immobilized enzyme was used for the decomposition of hydrogen peroxide in milk samples and also by using a catalase biosensor prepared the decomposition level of hydrogen peroxide was detected. PMID:15508280
Yildiz, Hatice; Akyilmaz, Erol; Din�kaya, Erhan
Heterogeneity of Catalase in Maturing and Germinated Cotton Seeds 1
To investigate possible charge and size heterogeneity of catalase (EC 18.104.22.168) in cotton (Gossypium hirsutum L. cv Deltapine 62), extracts of cotyledons from different developmental ages were subjected to nondenaturing polyacrylamide gel electrophoresis and isoelectric focusing. Special precautions (e.g. fresh homogenates, reducing media) were necessary to prevent artefacts due to enzyme modification during extraction and storage. When the gels were stained for enzyme activity, two distinct electrophoretic forms of catalase were resolved in extracts of maturing and mature cotton seeds. In germinated seeds, three additional cathodic forms were detected revealing a total of five electrophoretic variants. In green cotyledons, the two anodic forms characteristic of ungerminated seeds were less active; whereas, the most cathodic form was predominant. All forms of catalase were found in isolated glyoxysomes. Corresponding electrophoretic patterns were found on Western blots probed with anticatalase serum; no immunoreactive, catalytically inactive forms were detected. Western blots of sodium dodecyl sulfate-polyacrylamide gels revealed only one immunoreactive (55 kilodaltons) polypeptide in cotton extracts of all developmental ages. Results from isoelectric focusing and Ferguson plots indicate that the electrophoretic variants of catalase are charge isomers with a molecular weight of approximately 230,000. Images Fig. 1 Fig. 2 Fig. 3 Fig. 6 Fig. 7
Kunce, Christine M.; Trelease, Richard N.
We previously reported up-regulation of aryl hydrocarbon receptor (AhR) expression as a mechanism by which overexpression of Cu/Zn-superoxide dismutase (SOD) and/or catalase accelerates benzo(a)pyrene (BaP) detoxification in mouse aorta endothelial cells (MAECs). The objective of this study was to investigate the regulatory role of specificity protein-1 (Sp1) in AhR expression in MAECs that overexpress Cu/Zn-SOD and/or catalase. Our data demonstrated comparable levels of nuclear Sp1 protein in the transgenic and wild-type MAECs; however, binding of Sp1 protein to the AhR promoter region was more than 2-fold higher in MAECs overexpressing Cu/Zn-SOD and/or catalase than in wild-type cells. Inhibition of Sp1 binding to the AhR promoter by mithramycin A reduced AhR expression and eliminated the differences between wild-type MAECs, and three lines of transgenic cells. Functional promoter analysis indicated that AhR promoter activity was significantly higher in MAECs overexpressing catalase than in wild-type cells. Mutation of an AhR promoter Sp1-binding site or addition of hydrogen peroxide to the culture medium reduced AhR promoter activity, and decreased the differences between wild-type MAECs and transgenic cells overexpressing catalase. These results suggest that increased Sp1 binding to the AhR promoter region is an underlying mechanism for up-regulation of AhR expression in MAECs that overexpress Cu/Zn-SOD and/or catalase.
Tang, Tian; Lin, Xinghua; Yang, Hong; Zhou, LiChun; Wang, Zefen; Shan, Guang; Guo, ZhongMao
Catalase has only a minor role in protection against near-ultraviolet radiation damage in bacteria
In bacterial cells near-ultraviolet radiation (NUV) generates H2O2 which can be decomposed by endogenous catalase to H2O and O2. To assess the roles of H2O2 and catalase in NUV lethality, we manipulated the amount of intracellular catalase (a) by the use of mutant and plasmid strains with altered endogenous catalase, (b) physiologically, by the addition of glucose, and (c) by
A. Eisenstark; G. Perrot
Stimulation of catalase activity in carp ( Cyprinus�carpio ) ovarian follicles by gonadotropin in�vitro
The changes in catalase activity in carp (Cyprinus�carpio) ovarian follicles at different developmental stages were investigated. The effect of follicle-stimulating hormone (FSH) on catalase activity and estradiol secretion by carp ovarian follicles was studied to establish a developmental role of catalase in folliculogenesis in fish ovary. The follicular homogenates from large follicles showed higher (9.45��0.64�units\\/mg protein) catalase-specific activity than the
Rahul Behl
After whole-body exposure of 1000 r x rays, liver catalase activity of ; mlce was decreased for several days. The -catalase activity was decreased also ; after intraperttoneal administration of liver extract, urine, and serum prepared ; from the animal irradiated by x rays. Therefore, lt is suggested that the ; substance inhibiting liver catalase of mice is contained in
T. Yanagisawa; N. Hiramatsu; Z. Iwasaki; H. Toda
Cloning and Sequencing of a Candida albicans Catalase Gene and Effects of Disruption of This Gene
Catalase plays a key role as an antioxidant, protecting aerobic organisms from the toxic effects of hydrogen peroxide, and in some cases has been postulated to be a virulence factor. To help elucidate the function of catalase in Candida albicans, a single C. albicans-derived catalase gene, designated CAT1, was isolated and cloned. Degenerate PCR primers based on highly conserved areas
Comparative immunological study of catalases in the genus Micrococcus.
Double immunodiffusion tests were performed with crude extracts from various Micrococcus species and antisera against catalase of Micrococcus luteus CCM 169. Cell-free extracts of M. lylae ATCC 27566 exhibited good cross-reaction. Cell-free extracts or catalase enriched preparations of M. varians reacted very weakly and no reaction has been found with preparation of M. kristinae, M. nishinomiyaensis, M. roseus and M. sedentarius. The quantitative microcomplement fixation assay also revealed a closer relationship between M. luteus and M. lylae than between M. luteus and M. varians. Strains of other Micrococcus species reacted in the microcomplement assay with M. luteus antiserum just a weakly as non-related strains, e.g. Staphylococcus aureus or Cellulomonas cartalyticum. PMID:71880
Rupprecht, M; Schleifer, K H
Inhibition of the catalase reaction of Photosystem II by anions
A new binding site for anions which inhibit the water oxidizing complex (WOC) of Photosystem II in spinach has been identified. Anions which bind to this site inhibit the flash-induced S2\\/S0 catalase reaction (2H2O2?2H2O+O2) of the WOC by displacing hydrogen peroxide. Using a mass spectrometer and gas permeable membrane to detect the 32O2 product, the yield and lifetime of the
Junichi Mano; Kunio Kawamoto; G. Charles Dismukes; Kozi Asada
Dual targeting of yeast catalase A to peroxisomes and mitochondria.
Yeast catalase A (Cta1p) contains two peroxisomal targeting signals (SSNSKF) localized at its C-terminus and within the N-terminal third of the protein, which both can target foreign proteins to peroxisomes. In the present study we demonstrated that Cta1p can also enter mitochondria, although the enzyme lacks a classical mitochondrial import sequence. Cta1p co-targeting was studied in a catalase A null mutant after growth on different carbon sources, and expression of a Cta1p-GFP (green fluorescent protein)-fusion protein or a Cta1p derivative containing either a c-Myc epitope (Cta1p(myc)) or a SKF-extended tag (Cta1p(myc-SKF)). Peroxisomal and mitochondrial co-import of catalase A were tested qualitatively by fluorescence microscopy and functional complementation of a Delta cta1 null mutation, and quantitatively by subcellular fractionation followed by Western blot analysis and enzyme activity assays. Efficient Cta1p import into peroxisomes was observed when cells were cultivated under peroxisome-inducing conditions (i.e. growth on oleate), whereas significant co-import of Cta1p-GFP into mitochondria occurred when cells were grown under respiratory conditions that favour oxygen stress and ROS (reactive oxygen species) accumulation within this organelle. In particular, when cells were grown on the non-fermentable carbon source raffinose, respiration is maximally enhanced, and catalase A was efficiently targeted to the mitochondrial matrix where it presumably functions as scavenger of H2O2 and mitochondrial-derived ROS.
Petrova, Ventsislava Y; Drescher, Diane; Kujumdzieva, Anna V; Schmitt, Manfred J
Tritium effect in peroxidation of ehtanol by liver catalase.
1. Simultaneous determination of the rate of appearance of 3H in water from [(1R)-1-3H1] ethanol and the rate of acetaldehyde formation in the presence of rat or ox liver catalase under conditions of steady-state generation of H2O2 allowed calculation of the 3H isotope effect. The mean value of 2.52 obtained for rat liver catalase at 37 degrees C and pH 6.3-7.7 was independent of both ethanol concentration and the rate of H2O2 generation over a wide range. At 25 degrees C a slightly lower mean value of 2.40 was obtained with the ox liver catalase. 2. Neither the product, acetaldehyde, nor 4-methylpyrazole influenced the two rates measured in the assay. 3. Relating the value obtained for the 3H isotope effect to a known value for the 2H isotope effect strongly supports the view that both values are close to the true isotope effect with the respective substituted compounds on the rate constant in the catalytic step involving scission of the C-H bond. 4. The constancy of the isotope effect under various conditions makes it possible to use it for interpretations in vivo. 5. It was established that beta-D-galactose dehydrogenase exhibits B-specificity towards the nicotinamide ring in NAD.
Damgaard, S E
Progeric effects of catalase inactivation in human cells
Peroxisomes generate hydrogen peroxide, a reactive oxygen species, as part of their normal metabolism. A number of pathological situations exist in which the organelle's capacity to degrade the potentially toxic oxidant is compromised. It is the peroxidase, catalase, which largely determines the functional antioxidant capacity of the organelle, and it is this enzyme that is affected in aging, in certain diseases, and in response to exposure to specific chemical agents. To more tightly control the enzymatic activity of peroxisomal catalase and carefully document the effects of its impaired action on human cells, we employed the inhibitor 3-amino-1,2,4-triazole. We show that by chronically reducing catalase activity to approximately 38% of normal, cells respond in a dramatic manner, displaying a cascade of accelerated aging reactions. Hydrogen peroxide and related reactive oxygen species are produced, protein and DNA are oxidatively damaged, import into peroxisomes and organelle biogenesis is corrupted, and matrix metalloproteinases are hyper-secreted from cells. In addition, mitochondria are functionally impaired, losing their ability to maintain a membrane potential and synthesize reactive oxygen species themselves. These latter results suggest an important redox-regulated connection between the two organelle systems, a topic of considerable interest for future study.
Koepke, Jay I.; Wood, Christopher S.; Terlecky, Laura J. [Department of Pharmacology, Wayne State University School of Medicine, 540 E. Canfield Avenue, Detroit, Michigan, 48201 (United States); Walton, Paul A. [Department of Anatomy and Cell Biology, University of Western Ontario, London, Ontario (Canada); Terlecky, Stanley R. [Department of Pharmacology, Wayne State University School of Medicine, 540 E. Canfield Avenue, Detroit, Michigan, 48201 (United States)], E-mail: email@example.com
Immobilization of catalase by entrapping in alginate beads and catalase biosensor preparation for the determination of hydrogen peroxide decomposition.
In this study, catalase enzyme was immobilized by entrapping in alginate beads in the presence of gelatin. In the optimization studies of the bioactive layer immobilized some parameters such as enzyme amount, alginate, gelatin, and crosslinking agent glutaraldehyde amount were determined as 700 U/mL, 2.0%, 18 mg/mL, and 5.0%, respectively. Effects of pH and temperature on the immobilization were also investigated. In the characterization studies of the immobilized enzyme storage and thermal stability experiments were done. The immobilized enzyme was used for the decomposition of hydrogen peroxide in milk samples and also by using a catalase biosensor prepared by the decomposition level of hydrogen peroxide was detected. PMID:15508281
G�renek, G�lseren; Akyilmaz, Erol; Din�kaya, Erhan
Structure�Function Relationships in Fungal Large-Subunit Catalases
Neurospora crassa has two large-subunit catalases, CAT-1 and CAT-3. CAT-1 is associated with non-growing cells and accumulates particularly in asexual spores; CAT-3 is associated with growing cells and is induced under different stress conditions. It is our interest to elucidate the structure-function relationships in large-subunit catalases. Here we have determined the CAT-3 crystal structure and compared it with the previously determined CAT-1 structure. Similar to CAT-1, CAT-3 hydrogen peroxide (H{sub 2}O{sub 2}) saturation kinetics exhibited two components, consistent with the existence of two active sites: one saturated in the millimolar range and the other in the molar range. In the CAT-1 structure, we found three interesting features related to its unusual kinetics: (a) a constriction in the channel that conveys H{sub 2}O{sub 2} to the active site; (b) a covalent bond between the tyrosine, which forms the fifth coordination bound to the iron of the heme, and a vicinal cysteine; (c) oxidation of the pyrrole ring III to form a cis-hydroxyl group in C5 and a cis-{gamma}-spirolactone in C6. The site of heme oxidation marks the starts of the central channel that communicates to the central cavity and the shortest way products can exit the active site. CAT-3 has a similar constriction in its major channel, which could function as a gating system regulated by the H{sub 2}O{sub 2} concentration before the gate. CAT-3 functional tyrosine is not covalently bonded, but has instead the electron relay mechanism described for the human catalase to divert electrons from it. Pyrrole ring III in CAT-3 is not oxidized as it is in other large-subunit catalases whose structure has been determined. Different in CAT-3 from these enzymes is an occupied central cavity. Results presented here indicate that CAT-3 and CAT-1 enzymes represent a functional group of catalases with distinctive structural characteristics that determine similar kinetics.
Diaz, A.; Valdez, V; Rudino-Pinera, E; Horjales, E; Hansberg, W
Structure-function relationships in fungal large-subunit catalases.
Neurospora crassa has two large-subunit catalases, CAT-1 and CAT-3. CAT-1 is associated with non-growing cells and accumulates particularly in asexual spores; CAT-3 is associated with growing cells and is induced under different stress conditions. It is our interest to elucidate the structure-function relationships in large-subunit catalases. Here we have determined the CAT-3 crystal structure and compared it with the previously determined CAT-1 structure. Similar to CAT-1, CAT-3 hydrogen peroxide (H(2)O(2)) saturation kinetics exhibited two components, consistent with the existence of two active sites: one saturated in the millimolar range and the other in the molar range. In the CAT-1 structure, we found three interesting features related to its unusual kinetics: (a) a constriction in the channel that conveys H(2)O(2) to the active site; (b) a covalent bond between the tyrosine, which forms the fifth coordination bound to the iron of the heme, and a vicinal cysteine; (c) oxidation of the pyrrole ring III to form a cis-hydroxyl group in C5 and a cis-gamma-spirolactone in C6. The site of heme oxidation marks the starts of the central channel that communicates to the central cavity and the shortest way products can exit the active site. CAT-3 has a similar constriction in its major channel, which could function as a gating system regulated by the H(2)O(2) concentration before the gate. CAT-3 functional tyrosine is not covalently bonded, but has instead the electron relay mechanism described for the human catalase to divert electrons from it. Pyrrole ring III in CAT-3 is not oxidized as it is in other large-subunit catalases whose structure has been determined. Different in CAT-3 from these enzymes is an occupied central cavity. Results presented here indicate that CAT-3 and CAT-1 enzymes represent a functional group of catalases with distinctive structural characteristics that determine similar kinetics. PMID:19109972
D�az, Adelaida; Vald�s, V�ctor-Juli�n; Rudi�o-Pi�era, Enrique; Horjales, Eduardo; Hansberg, Wilhelm
Are catalase -844A/G polymorphism and activity associated with childhood obesity?
Catalase (CAT) is a peroxisomal antioxidant enzyme that is up-regulated upon oxidative stress. Previous studies have found associations between some single nucleotide polymorphisms (SNPs) located in the CAT promoter region in a variety of metabolic diseases. This is the first study that analyzes the association between erythrocyte CAT activity and candidate CAT SNPs with childhood obesity. The association study showed a significant positive association of the promoter variant -844A/G with childhood obesity and biomarkers of obesity such as weight, body mass index (BMI), BMI Z-Score, and adipocyte fatty acid-binding protein, along with a tendency toward significance with insulin resistance biomarkers. In addition, CAT erythrocyte activity was found to be significantly lower in obese children, and it was significantly correlated with obesity and insulin resistance biomarkers. No association was found between erythrocyte CAT activity and the SNP -844A/G. However, further in vitro and in vivo studies are needed to fully understand the role of CAT activity and SNPs in the development of insulin resistance in the setting of obesity. We hypothesize that CAT plays a role in early metabolic complications of obesity. PMID:23641975
Rup�rez, Azahara I; Olza, Josune; Gil-Campos, Mercedes; Leis, Rosaura; Mesa, Mar�a D; Tojo, Rafael; Ca�ete, Ram�n; Gil, Angel; Aguilera, Concepci�n M
Unique Presence of a Manganese Catalase in a Hyperthermophilic Archaeon, Pyrobaculum calidifontis VA1
We had previously isolated a facultatively anaerobic hyperthermophilic archaeon, Pyrobaculum calidifontis strain VA1. Here, we found that strain VA1, when grown under aerobic conditions, harbors high catalase activity. The catalase was purified 91-fold from crude extracts and displayed a specific activity of 23,500 U/mg at 70�C. The enzyme exhibited a Km value of 170 mM toward H2O2 and a kcat value of 2.9 � 104 s?1�subunit?1 at 25�C. Gel filtration chromatography indicated that the enzyme was a homotetramer with a subunit molecular mass of 33,450 Da. The purified catalase did not display the Soret band, which is an absorption band particular to heme enzymes. In contrast to typical heme catalases, the catalase was not strongly inhibited by sodium azide. Furthermore, with plasma emission spectroscopy, we found that the catalase did not contain iron but instead contained manganese. Our biochemical results indicated that the purified catalase was not a heme catalase but a manganese (nonheme) catalase, the first example in archaea. Intracellular catalase activity decreased when cells were grown anaerobically, while under aerobic conditions, an increase in activity was observed with the removal of thiosulfate from the medium, or addition of manganese. Based on the N-terminal amino acid sequence of the purified protein, we cloned and sequenced the catalase gene (katPc). The deduced amino acid sequence showed similarity with that of the manganese catalase from a thermophilic bacterium, Thermus sp. YS 8-13. Interestingly, in the complete archaeal genome sequences, no open reading frame has been assigned as a manganese catalase gene. Moreover, a homology search with the sequence of katPc revealed that no orthologue genes were present on the archaeal genomes, including those from the �aerobic� (hyper)thermophilic archaea Aeropyrum pernix, Sulfolobus solfataricus, and Sulfolobus tokodaii. Therefore, KatPc can be considered a rare example of a manganese catalase from archaea.
Amo, Taku; Atomi, Haruyuki; Imanaka, Tadayuki
Purification and Characterization of Catalase from Marine Bacterium Acinetobacter sp. YS0810
The catalase from marine bacterium Acinetobacter sp. YS0810 (YS0810CAT) was purified and characterized. Consecutive steps were used to achieve the purified enzyme as follows: ethanol precipitation, DEAE Sepharose ion exchange, Superdex 200 gel filtration, and Resource Q ion exchange. The active enzyme consisted of four identical subunits of 57.256?kDa. It showed a Soret peak at 405?nm, indicating the presence of iron protoporphyrin IX. The catalase was not apparently reduced by sodium dithionite but was inhibited by 3-amino-1,2,4-triazole, hydroxylamine hydrochloride, and sodium azide. Peroxidase-like activity was not found with the substrate o-phenylenediamine. So the catalase was determined to be a monofunctional catalase. N-terminal amino acid of the catalase analysis gave the sequence SQDPKKCPVTHLTTE, which showed high degree of homology with those of known catalases from bacteria. The analysis of amino acid sequence of the purified catalase by matrix-assisted laser desorption ionization time-of-flight mass spectrometry showed that it was a new catalase, in spite of its high homology with those of known catalases from other bacteria. The catalase showed high alkali stability and thermostability.
Fu, Xinhua; Wang, Wei; Hao, Jianhua; Zhu, Xianglin; Sun, Mi
Catalase degradation in sunflower cotyledons during peroxisome transition from glyoxysomal to leaf peroxisomal function. [Helianthus annuus
First order rate constant for the degradation (degradation constants) of catalase in the cotyledons of sunflower (Helianthus annuus L.) were determined by measuring the loss of catalase containing /sup 14/C-labeled heme. During greening of the cotyledons, a period when peroxisomes change from glyoxysomal to leaf peroxisomal function, the degradation of glyoxysomal catalase is significantly slower than during all other stages of cotyledon development in light or darkness. The degradation constant during the transition stage of peroxisome function amounts to 0.205 day/sup -1/ in contrast to the constants ranging from 0.304 day/sup -1/ to 0.515 day/sup -1/ during the other developmental stages. Density labeling experiments comprising labeling of catalase with /sup 2/H/sub 2/O and its isopycnic centrifugation on CsCl gradients demonstrated that the determinations of the degradation constants were not substantially affected by reutilization of /sup 14/C-labeled compounds for catalase synthesis. The degradation constants for both glyoxysomal catalase and catalase synthesized during the transition of peroxisome function do not differ. This was shown by labeling the catalases with different isotopes and measuring the isotope ratio during the development of the cotyledons. The results are inconsistent with the concept that an accelerated and selective degradation of glyoxysomes underlies the change in peroxisome function. The data suggest that catalase degradation is at least partially due to an individual turnover of catalase and does not only result from a turnover of the whole peroxisomes.
Eising, R.; Gerhardt, B.
Wood Utilization Is Dependent on Catalase Activities in the Filamentous Fungus Podospora anserina
Catalases are enzymes that play critical roles in protecting cells against the toxic effects of hydrogen peroxide. They are implicated in various physiological and pathological conditions but some of their functions remain unclear. In order to decipher the role(s) of catalases during the life cycle of Podospora anserina, we analyzed the role of the four monofunctional catalases and one bifunctional catalase-peroxidase genes present in its genome. The five genes were deleted and the phenotypes of each single and all multiple mutants were investigated. Intriguingly, although the genes are differently expressed during the life cycle, catalase activity is dispensable during both vegetative growth and sexual reproduction in laboratory conditions. Catalases are also not essential for cellulose or fatty acid assimilation. In contrast, they are strictly required for efficient utilization of more complex biomass like wood shavings by allowing growth in the presence of lignin. The secreted CATB and cytosolic CAT2 are the major catalases implicated in peroxide resistance, while CAT2 is the major player during complex biomass assimilation. Our results suggest that P. anserina produces external H2O2 to assimilate complex biomass and that catalases are necessary to protect the cells during this process. In addition, the phenotypes of strains lacking only one catalase gene suggest that a decrease of catalase activity improves the capacity of the fungus to degrade complex biomass.
Bourdais, Anne; Bidard, Frederique; Zickler, Denise; Berteaux-Lecellier, Veronique; Silar, Philippe; Espagne, Eric
Nanoparticle-mediated catalase delivery protects human neurons from oxidative stress
Several neurodegenerative diseases and brain injury involve reactive oxygen species and implicate oxidative stress in disease mechanisms. Hydrogen peroxide (H2O2) formation due to mitochondrial superoxide leakage perpetuates oxidative stress in neuronal injury. Catalase, an H2O2-degrading enzyme, thus remains an important antioxidant therapy target. However, catalase therapy is restricted by its labile nature and inadequate delivery. Here, a nanotechnology approach was evaluated using catalase-loaded, poly(lactic co-glycolic acid) nanoparticles (NPs) in human neuronal protection against oxidative damage. This study showed highly efficient catalase encapsulation capable of retaining?99% enzymatic activity. NPs released catalase rapidly, and antioxidant activity was sustained for over a month. NP uptake in human neurons was rapid and nontoxic. Although human neurons were highly sensitive to H2O2, NP-mediated catalase delivery successfully protected cultured neurons from H2O2-induced oxidative stress. Catalase-loaded NPs significantly reduced H2O2-induced protein oxidation, DNA damage, mitochondrial membrane transition pore opening and loss of cell membrane integrity and restored neuronal morphology, neurite network and microtubule-associated protein-2 levels. Further, catalase-loaded NPs improved neuronal recovery from H2O2 pre-exposure better than free catalase, suggesting possible applications in ameliorating stroke-relevant oxidative stress. Brain targeting of catalase-loaded NPs may find wide therapeutic applications for oxidative stress-associated acute and chronic neurodegenerative disorders.
Singhal, A; Morris, V B; Labhasetwar, V; Ghorpade, A
Neuroprotective effect of adenoviral catalase gene transfer in cortical neuronal cultures
Reduced availability of reactive oxygen species is a key component of neuroprotection against various toxic stimuli. Recently we showed that the hydrogen peroxide scavenger catalase plays a central role in delayed preconditioning induced by the mitochondrial ATP-sensitive potassium channel opener BMS-191095. The purpose of the experiments discussed here was to investigate the neuroprotective effect of catalase in vitro using a recombinant adenoviral catalase gene transfer protocol. To induce catalase overexpression, cultured rat cortical neurons were infected with the adenoviral vector Ad5CMVcatalase and control cells were incubated with Ad5CMVntLacZ for 24h. Gene transfer effectively increased catalase protein levels and activity, but did not influence other antioxidants tested. Ad5CMVcatalase, with up to 10 plaque forming units (pfu) per neuron, did not affect cell viability under control conditions and did not protect against glutamate excitotoxicity or oxygen-glucose deprivation. In contrast, catalase overexpression conferred a dose-dependent protection against exposure to hydrogen peroxide (viability: control, 33.02�1.09%; LacZ 10 pfu/cell, 32.85�1.51%; catalase 1 pfu/cell, 62.09�4.17%*; catalase 2 pfu/cell, 98.71�3.35%*; catalase 10 pfu/cell, 99.68�1.99%*; *p<0.05 vs. control; mean�SEM). Finally, the protection could be antagonized using the catalase inhibitor 3-aminotriazole. Our results support the view that enhancing cellular antioxidant capacity may play a crucial role in neuroprotective strategies.
Gaspar, Tamas; Domoki, Ferenc; Lenti, Laura; Institoris, Adam; Snipes, James A; Bari, Ferenc; Busija, David W
Plating isolation of various catalase-negative microorganisms from soil
NASA Technical Reports Server (NTRS)
A unique plating procedure was developed that allows isolation, but not enumeration, of representatives of the catalase-negative soil microflora. The numbers recovered, however, are low as compared to the numbers recovered when the modified dilution-to-extinction isolation procedure is used. The latter procedure provides prolonged inoculation in sealed tubes containing a nutritionally rich broth medium over small submerged agar slants. In contrast, the plating procedure utilizes nutritionally minimal media and the shorter incubations mandated by the inherent problems associated with plating.
Labeda, D. P.; Hunt, C. M.; Casida, L. E., Jr.
White, David C. (The Rockefeller Institute, New York, N. Y.) Cytochrome and catalase patterns during growth of Haemophilus parainfluenzae. J. Bacteriol. 83:851�859. 1962.�By following the cytochrome and catalase concentrations during the growth cycle and under various growth conditions in Haemophilus parainfluenzae, a rapid increase in the cytochrome oxidases and a large increase in cytochrome c1 concentration can be demonstrated between log-phase and stationary-phase cells and between vigorously aerated and anaerobic growth conditions. The three cytochrome oxidases develop differentially under various growth conditions. The principal oxidase formed in vigorously aerated cultures is cytochrome o. With limited aeration, maximal development of cytochrome a2 occurs; with anaerobically grown cells, there is a marked increase in the concentration of cytochrome a1. With the rapid increase in cytochrome c1 concentration, soluble, nonenzymatically reducible cytochrome c1 is also formed, which remains in the bacterial cell sap. From these data it is postulated that the electron-transport system is assembled from individual components which can be modified by the growth conditions. The cytochrome c1 may be synthesized in the cell sap and then incorporated into the electron-transport system.
White, David C.
Extension of mouse lifespan by overexpression of catalase.
The free radical theory of aging was originally proposed 50 years ago, and is arguably the most popular mechanism explaining the aging process. According to this theory, aging results from the progressive decline in organ function due to the damage generated by reactive oxygen species (ROS). These chemical species are a normal part of metabolism, and a group of enzymes exists to protect cells against their toxic effects. One of these species is hydrogen peroxide (H(2)O(2)), which can be degraded by catalase. To determine the role of hydrogen peroxide in aging and its importance in different subcellular compartments, transgenic mice were developed with increased catalase activities localized to the peroxisome (PCAT), nucleus (NCAT), or mitochondrion (MCAT). The largest effect on lifespan was found in MCAT animals, with a 20% increase in median lifespan and a 10% increase in the maximum lifespan. A more modest effect was seen in PCAT animals, and no significant change was found in NCAT animals. Upon further examination of the MCAT mice, it was found that H(2)O(2) production and H(2)O(2)-induced aconitase inactivation were attenuated, oxidative damage and the development of mitochondrial deletions were reduced, and cardiac pathology and cataract development were delayed. These results are consistent with a role of H(2)O(2) in the development of pathology and in the limitation of mouse lifespan. They also demonstrate the importance of mitochondria as a source, and possible target, of ROS. PMID:19943142
Schriner, Samuel E; Linford, Nancy J
A gasometric method to determine erythrocyte catalase activity.
We describe a new gasometric method to determine erythrocyte catalase activity by the measurement of the volume of oxygen produced as a result of hydrogen peroxide decomposition in a system where enzyme and substrate are separated in a special reaction test tube connected to a manometer and the reagents are mixed with a motor-driven stirrer. The position of the reagents in the test tube permits the continuous measurement of oxygen evolution from the time of mixing, without the need to stop the reaction by the addition of acid after each incubation time. The enzyme activity is reported as K Hb, i.e., mg hydrogen peroxide decomposed per second per gram of hemoglobin (s-1 g Hb-1). The value obtained for catalase activity in 28 samples of hemolyzed human blood was 94.4 +/- 6.17 mg H2O2 s-1 g Hb-1. The results obtained were precise and consistent, indicating that this rapid, simple and inexpensive method could be useful for research and routine work. PMID:10464384
Siqueira, A J; Remi�o, J O; Azevedo, A M; Azambuja, C R
A method for molecular analysis of catalase gene diversity in seawater.
Catalase plays an important role in the metabolism of marine bacteria and has potential impact on the marine environment. Four PCR primers were designed to amplify the catalase gene fragments in marine bacteria by applying metagenomic DNA from Yellow Sea surface water as the template. Of the four reproducible target PCR products, the longest one with 900�bp were chosen for catalase gene library construction by the T-vector and the white Escherichia coli colonies in the library was screened through restriction-digesting the reamplified insert fragments by the selected restriction endonuclease MboI, and then the bands of the resulting products were displayed in the agarose gel by electrophoresis. The unique restriction fragment length polymorphism (RFLP) pattern was selected and the corresponding catalase gene fragments were sequenced, which verified that every unique RFLP pattern represented one type of catalase. This PCR-RFLP method above was established to investigate the bacterial catalase diversity in seawater. PMID:24426153
Wang, Wei; Ji, Xiaofeng; Yuan, Cui; Dai, Fangqun; Zhu, Jiancheng; Sun, Mi
Rapid upregulation of heart antioxidant enzymes during arousal from estivation in the Giant African snail (Achatina fulica).
Estivation is an adaptive response to environments characterized by elevated temperatures and desiccative stress, as may occur during summer dry seasons. Similar to diapause and hibernation, it is characterized by low levels of activity, a drastically suppressed metabolic rate and enhanced stress resistance. We tested the hypothesis that Achatina fulica, a pulmonate land snail, enhances stress resistance during estivation and/or arousal by upregulating intracellular antioxidant defenses in the heart, kidney, hepatopancreas and foot tissues. No statistically significant changes in mitochondrial or cytosolic superoxide dismutase levels or activities, or glutathione peroxidase, glutathione reductase or catalase activities were associated with estivation in any tissue, however. In contrast, during arousal from estivation, activities of several antioxidant enzymes increased in heart, hepatopancreas and foot. In heart, a rapid increase in MnSOD protein levels was observed that peaked at 2h post arousal, but no such change was observed in CuZnSOD protein levels. Glutathione peroxidase activity was upregulated at 1h post arousal and remained elevated until 8h post arousal in heart tissue. Glutathione peroxidase was also upregulated at 24h post arousal in foot tissue. Glutathione reductase activity was upregulated at 4h post arousal in heart and foot tissues whereas catalase activity showed no changes. Markers of lipid peroxidation and protein damage revealed no significant increases during estivation or arousal. Therefore, antioxidant enzymes may play a role in oxidative stress defense specifically during arousal from estivation in A. fulica. PMID:20621194
Salway, Kurtis D; Tattersall, Glenn J; Stuart, Jeffrey A
Distribution of oxidase enzymes in potato tubers relative to blackspot susceptibility II. Peroxidase and Catalase
Peroxidase and catalase activities in selected potato tuber tissue were studied for differences and possible association with\\u000a resistance or susceptibility to blackspot. Unbruised stem-end tissue had significantly greater peroxidase activity than did\\u000a unbruised bud-end tissue. However, there was no significant difference between catalase activity at either end of the tuber;\\u000a between blackspot susceptibility and peroxidase or catalase activity; and between
M. L. Weaver; E. Hautala; R. M. Reeve
Impedance spectroscopy and conductometric biosensing for probing catalase reaction with cyanide as ligand and inhibitor
In this work, a new biosensor was prepared through immobilization of bovine liver catalase in a photoreticulated poly (vinyl alcohol) membrane at the surface of a conductometric transducer. This biosensor was used to study the kinetics of catalase�H202 reaction and its inhibition by cyanide. Immobilized catalase exhibited a Michaelis�Menten behaviour at low H202 concentrations (<100mM) with apparent constant KMapp=84�3mM and
Naima Bouyahia; Mohamed Larbi Hamlaoui; Mouna Hnaien; Florence Lagarde; Nicole Jaffrezic-Renault
Catalase A from Botrytis cinerea is not expressed during infection on tomato leaves
Catalase mediates the enzymatic breakdown of hydrogen peroxide to water and molecular oxygen. During the infection of broad bean (Vicia faba) byBotrytis cinereathe release of hydrogen peroxide by fungal oxidase activity was proposed to facilitate penetration by the pathogen. Catalase activity might play a role in protecting the fungus against the damaging effects of hydrogen peroxide.A cDNA clone encoding catalase
C. J. B. van der Vlugt-Bergmans; C. A. M. Wagemakers; D. C. T. Dees; J. A. L. van Kan
Comparative analysis of catalases: spectral evidence against heme-bound water for the solution enzymes
A recent X-ray structural analysis of M. luteus catalase indicates heme-bound H2O trans to the proximal tyrosinate ligand, a finding in contrast to previous X-ray data reporting a 5-coordinate heme for bovine liver catalase. The presence of heme-bound H2O, requiring displacement prior to substrate-binding, is likely to be catalytically significant for catalases. We have used magnetic circular dichroism (MCD) spectroscopy,
Laura A. Andersson; Anna K. Johnson; Melissa D. Simms; Timothy R. Willingham
Catalase-Peroxidase from Synechocystis Is Capable of Chlorination and Bromination Reactions
Catalase-peroxidases (KatGs) are multifunctional heme peroxidases exhibiting an overwhelming catalase activity and a substantial peroxidase activity of broad specificity. Here, we show that catalase-peroxidases are also haloperoxidases capable of oxidizing chloride, bromide, and iodide in a peroxide- and enzyme-dependent manner. Recombinant KatG and the variants R119A, W122F, and W122A from the cyanobacterium Synechocystis PCC 6803 have been tested for their
Christa Jakopitsch; G�nther Regelsberger; Paul Georg Furtm�ller; Florian R�ker; G�nter A Peschek; Christian Obinger
Regulation of catalase activity in leaves of Nicotiana sylvestris by high CO sub 2
The effect of high CO{sub 2} (1% CO{sub 2}/21% O{sub 2}) on the activity of specific forms of catalase (CAT-1, -2, and -3) in seedling leaves of tobacco (Nicotiana sylvestris, Nicotiana tabacum) was examined. In high CO{sub 2} total catalase activity decreased by 50% in the first 2 days, followed by a more gradual decline in the next 4 days. The loss of total activity resulted primarily from a decrease in CAT-1 catalase. In contrast, the activity of CAT-3 catalase, a form with enhanced peroxidatic activity, increased 3-fold in high CO{sub 2} relative to air controls after 4 days. Short-term exposure to high CO{sub 2} indicated that the 50% loss of total activity occurs in the firs 12 hours. Catalase levels increased to normal within 12 hours after seedlings were returned to air. When seedlings were transferred to air after prolonged exposure to high CO{sub 2} (13 days), the levels of CAT-1 catalase were partially restored while CAT-3 remained at its elevated level. Levels of superoxide dismutase activity and those of several peroxisomal enzymes were not affected by high CO{sub 2}. Total catalase levels did not decline when seedlings were exposed to atmospheres of 0.04% CO{sub 2}/5% O{sub 2} or 0.04% CO{sub 2}/1% O{sub 2}, indicating that regulation of catalase in high CO{sub 2} is not related directly to suppression of photorespiration. Antibodies prepared against CAT-1 catalase from N. tabacum reacted strongly against CAT-1 catalase from both N. sylvestris and N. tabacum but not against CAT-3 catalase from either species.
Havir, E.A.; McHale, N.A. (Connecticut Agricultural Experiment Station, New Haven (USA))
Study of catalase adsorption on two mixed-mode ligands and the mechanism involved therein.
Mixed-mode chromatography sorbents n-hexylamine HyperCel� (HEA) and phenylpropylamine HyperCel� (PPA) were evaluated for the study of adsorption of catalase from two different sources. Various parameters such as buffer composition, ionic strength and pH were investigated to study the mechanism of interaction of commercially available pre-purified catalase from Bovine liver, purified catalase from black gram (Vigna mungo) and crude extract of black gram containing catalase with these mixed-mode ligands. A simple and economical screening protocol for identifying optimal buffer conditions for adsorption and desorption of catalase was established with micro volumes of the sorbent in batch mode. With HEA HyperCel, it was observed that pre-purified catalase from both bovine liver and black gram was completely retained at pH 7.0, irrespective of the presence or absence of NaCl in the adsorption buffer, whereas the catalase from crude extract of black gram was completely retained only in the presence of 0.2 M salt in the adsorption buffer. The elution of catalase from both the sources was accomplished by lowering the pH to 4.5 in absence of salt. In case of PPA HyperCel, catalase from both the sources was very strongly adsorbed under different buffer conditions studied, and elution did not yield a significant catalase activity. From the screening experiments, it could be concluded that the interaction of catalase with HEA HyperCel could be dominated by hydrophobic forces with minor contributions from ionic interaction and with PPA HyperCel, it could be a combination of different non-covalent interactions acting on different loci on the surface of the protein. PMID:23108613
Shiva Ranjini, S; Vijayalakshmi, M A
Regulation of Catalase Activity in Leaves of Nicotiana sylvestris by High CO2
The effect of high CO2 (1% CO2/21% O2) on the activity of specific forms of catalase (CAT-1, -2, and -3) (EA Havir, NA McHale [1987] Plant Physiol 84: 450-455) in seedling leaves of tobacco (Nicotiana sylvestris, Nicotlana tabacum) was examined. In high CO2, total catalase activity decreased by 50% in the first 2 days, followed by a more gradual decline in the next 4 days. The loss of total activity resulted primarily from a decrease in CAT-1 catalase. In contrast, the activity of CAT-3 catalase, a form with enhanced peroxidatic activity, increased 3-fold in high CO2 relative to air controls after 4 days. Short-term exposure to high CO2 indicated that the 50% loss of total activity occurs in the first 12 hours. Catalase levels increased to normal within 12 hours after seedlings were returned to air. When seedlings were transferred to air after prolonged exposure to high CO2 (13 days), the levels of CAT-1 catalase were partially restored while CAT-3 remained at its elevated level. Levels of superoxide dismutase activity and those of several peroxisomal enzymes were not affected by high CO2. Total catalase levels did not decline when seedlings were exposed to atmospheres of 0.04% CO2/5% O2 or 0.04% CO2/1% O2, indicating that regulation of catalase in high CO2 is not related directly to suppression of photorespiration. Antibodies prepared against CAT-1 catalase from N. tabacum reacted strongly against CAT-1 catalase from both N. sylvestris and N. tabacum but not against CAT-3 catalase from either species. This observation, along with the rapid changes in CAT-1 and the much slower changes in CAT-3 suggest that one form is not directly derived from the other. Images Figure 4
Havir, Evelyn A.; McHale, Neil A.
Effect of a gas environment on catalase cryolysis
NASA Technical Reports Server (NTRS)
Catalase is inactivated during repeated freezing and thawing of its solutions. The effect depends on the temperature of freezing and the gas atmosphere. Gases tested for the degree of their effect on cryolysis in order of increasing effect on inactivation of the enzymes are: N2, He, O2, and H2. In a gas environment consisting of oxygen and nitrogen an additive summation of the effects on cryolysis which were produced by each gas separately is observed. The effect of an atmosphere consisting of hydrogen and argon and cryolysis of an enzyme revealed a significant increase of inactivation in comparison to the expected motivation for the case of additive summation of effects produced by each gas separately.
Komolova, G. S.
The catalase-hydrogen peroxide system. Role of sub-units in the thermal deactivation of bacterial catalase in the absence of substrate
1. Kinetic studies of the thermal deactivation of bacterial catalase in the absence of substrate suggest that the reaction involves a protonation-induced reversible dissociation of catalase into catalatically inactive sub-units, followed by an irreversible transformation of the sub-units into deactivated products. It is possible that the sub-units are mono-haem species. The rate of deactivation decreases with increasing pressure in accordance with the predictions of the proposed model. 2. The results also imply that the addition of hydrogen peroxide substrate induces the re-formation of active catalase. Under appropriate conditions the activity of catalase is found to increase with time in a manner that is quantitatively consistent with the results of deactivation studies.
Jones, Peter; Suggett, A.
The molecular characterization of a catalase from Chinese mitten crab Eriocheir sinensis.
Catalase (CAT) is an antioxidant enzyme and plays a significant role in the protection against oxidative stress by reducing hydrogen peroxide. The CAT cDNA of Eriocheir sinensis (EsCAT) was cloned via RACE technique. The complete sequence of EsCAT cDNA consisted of a 5' untranslated regions (UTR) of 224 bp, a 3' UTR of 1287 bp with a poly (A) tail and an open reading frame (ORF) of 1542 bp, which encoded a polypeptide of 513 amino acid residues with a calculated molecular mass of approximately 58.86 kDa and a theoretical isoelectric point of 6.880. The deduced amino acid sequence of EsCAT contained a highly conserved proximal active-site signature motif ((60)FDRERIPERVVHAKGAL(76)) and a proximal heme-ligand signature motif ((350)RLFSYNDTH(358)) and exhibited high similarity with other reported CATs. In the phylogenetic tree, EsCAT was clustered with the CATs from Scylla serrata and Portunus trituberculatus. The EsCAT transcripts were constitutively expressed in haepatopancreas, haemocytes, gill, gonad, muscle and heart, with highest expression level in haepatopancreas. The relative expression level of EsCAT mRNA in haemocytes was continuously up-regulated and reached the peak level at 48 h post-Vibrio anguillarum challenge. The purified recombinant EsCAT protein displayed antioxidant activity against hydrogen peroxide with high thermal stability and broad spectrum of pH values. All these results demonstrated that EsCAT was an efficient antioxidant enzyme and potentially involved in the regulation of redox and innate immune response of crabs. PMID:23171400
Wang, M; Wang, L; Zhou, Z; Gao, Y; Wang, L; Shi, X; Gai, Y; Mu, C; Song, L
A Laboratory Experiment Investigating Different Aspects of Catalase Activity in an Inquiry - Based Approach
The action of the enzyme catalase on aqueous hydrogen peroxide to generate oxygen gas is a well-established demonstration (1-3). Catalase is typically obtained by aqueous extraction of a potato, and the potato extract is mixed together with 3% hydrogen peroxide. The oxygen that is produced can be collected over water. Variations on the procedure can demonstrate the dependence of catalytic
Doris R. Kimbrough; Mary Ann Magoun; Meg Langfur
Activation?Based Catalase Enzyme Electrode and its Usage for Glucose Determination
In this study, a novel type amperometric biosensor, which is based on the activation of catalase enzyme by glucose, was developed and used for the sensitive determination of glucose. For the preparation of the biosensor catalase enzyme was immobilized in gelatin by using cross?linking agent glutaraldehyde and fixed on a pretreated teflon membrane of a dissolved oxygen probe. Glucose was
Pinar Akbayirli; Erol Akyilmaz
Inhibition of Catalase by Tea Catechins in Free and Cellular State: A Biophysical Approach
Tea flavonoids bind to variety of enzymes and inhibit their activities. In the present study, binding and inhibition of catalase activity by catechins with respect to their structure-affinity relationship has been elucidated. Fluorimetrically determined binding constants for (?)-epigallocatechin gallate (EGCG) and (?)-epicatechin gallate (ECG) with catalase were observed to be 2.27�106 M?1 and 1.66�106 M?1, respectively. Thermodynamic parameters evidence exothermic and spontaneous interaction between catechins and catalase. Major forces of interaction are suggested to be through hydrogen bonding along with electrostatic contributions and conformational changes. Distinct loss of ?-helical structure of catalase by interaction with EGCG was captured in circular dichroism (CD) spectra. Gallated catechins demonstrated higher binding constants and inhibition efficacy than non-gallated catechins. EGCG exhibited maximum inhibition of pure catalase. It also inhibited cellular catalase in K562 cancer cells with significant increase in cellular ROS and suppression of cell viability (IC50 54.5 �M). These results decipher the molecular mechanism by which tea catechins interact with catalase and highlight the potential of gallated catechin like EGCG as an anticancer drug. EGCG may have other non-specific targets in the cell, but its anticancer property is mainly defined by ROS accumulation due to catalase inhibition.
Pal, Sandip; Dey, Subrata Kumar; Saha, Chabita
The in Vivo Toxicity of Hydroxyurea Depends on Its Direct Target Catalase*
Hydroxyurea (HU) is a well tolerated ribonucleotide reductase inhibitor effective in HIV, sickle cell disease, and blood cancer therapy. Despite a positive initial response, however, most treated cancers eventually progress due to development of HU resistance. Although RNR properties influence HU resistance in cell lines, the mechanisms underlying cancer HU resistance in vivo remain unclear. To address this issue, we screened for HU resistance in the plant Arabidopsis thaliana and identified seventeen unique catalase mutants, thereby establishing that HU toxicity depends on catalase in vivo. We further demonstrated that catalase is a direct HU target by showing that HU acts as a competitive inhibitor of catalase-mediated hydrogen peroxide decomposition. Considering also that catalase can accelerate HU decomposition in vitro and that co-treatment with another catalase inhibitor alleviates HU effects in vivo, our findings suggests that HU could act as a catalase-activated pro-drug. Clinically, we found high catalase activity in circulating cells from untreated chronic myeloid leukemia, offering a possible explanation for the efficacy of HU against this malignancy.
Juul, Trine; Malolepszy, Anna; Dybkaer, Karen; Kidmose, Rune; Rasmussen, Jan Trige; Andersen, Gregers Rom; Johnsen, Hans Erik; J?rgensen, Jan-Elo; Andersen, Stig Uggerh?j
The in vivo toxicity of hydroxyurea depends on its direct target catalase.
Hydroxyurea (HU) is a well tolerated ribonucleotide reductase inhibitor effective in HIV, sickle cell disease, and blood cancer therapy. Despite a positive initial response, however, most treated cancers eventually progress due to development of HU resistance. Although RNR properties influence HU resistance in cell lines, the mechanisms underlying cancer HU resistance in vivo remain unclear. To address this issue, we screened for HU resistance in the plant Arabidopsis thaliana and identified seventeen unique catalase mutants, thereby establishing that HU toxicity depends on catalase in vivo. We further demonstrated that catalase is a direct HU target by showing that HU acts as a competitive inhibitor of catalase-mediated hydrogen peroxide decomposition. Considering also that catalase can accelerate HU decomposition in vitro and that co-treatment with another catalase inhibitor alleviates HU effects in vivo, our findings suggests that HU could act as a catalase-activated pro-drug. Clinically, we found high catalase activity in circulating cells from untreated chronic myeloid leukemia, offering a possible explanation for the efficacy of HU against this malignancy. PMID:20452979
Juul, Trine; Malolepszy, Anna; Dybkaer, Karen; Kidmose, Rune; Rasmussen, Jan Trige; Andersen, Gregers Rom; Johnsen, Hans Erik; J�rgensen, Jan-Elo; Andersen, Stig Uggerh�j
Root-colonizing, saprophytic fluorescent pseudomonads of the Pseudomonas putida-P. fluorescens group express similar levels of catalase and superoxide dismutase activities during growth on a sucrose- and amino acid-rich medium. ncreased specific activities of catalase but not sup...
Psuedomonas putida, a saprophytic root-colonizing bacterium, produces multiple forms of catalase Catalase A which increases in specific activity during growth phase and after treatment with H2O2, is located in the and is inhibited by 3-amino-1,2-4-triazole, EDTA, and cyanide, but...
Encapsulation of catalase in polyelectrolyte microspheres composed of melamine formaldehyde, dextran sulfate, and protamine.
Immobilization of catalase (molecular weight 240,000 daltons) in polyelectrolyte microspheres was studied. The microspheres were obtained by alternating adsorption of dextran sulfate and protamine on commercially available melamine formaldehyde cores followed by the core hydrolysis at pH 1.7. As the interior of the microspheres was filled with homogeneous matrix, the catalase distribution inside the microspheres was uniform. The quantity of entrapped catalase was dependent on the initial concentration of the enzyme and pH of solution, and the peak value was 10(8)-10(9) molecules per microsphere. It was demonstrated that catalase was entrapped in the microspheres via electrostatic and hydrophobic interactions. The catalase activity inside the microspheres increased as the quantity of enzyme decreased, which was due to the switch between diffusion and kinetic regimes of the enzymatic reaction. The microspheres could be applied for separation and concentration of high molecular weight proteins. PMID:15310276
Balabushevich, N G; Zimina, E P; Larionova, N I
Catalase activity is modulated by calcium and calmodulin in detached mature leaves of sweet potato.
Catalase (CAT) functions as one of the key enzymes in the scavenging of reactive oxygen species and affects the H2O2 homeostasis in plants. In sweet potato, a major catalase isoform was detected, and total catalase activity showed the highest level in mature leaves (L3) compared to immature (L1) and completely yellow, senescent leaves (L5). The major catalase isoform as well as total enzymatic activity were strongly suppressed by ethylene glycol-bis(2-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA). This inhibition could be specifically and significantly mitigated in mature L3 leaves by exogenous CaCl2, but not MgCl2 or CoCl2. EGTA also inhibited the activity of the catalase isoform in vitro. Furthermore, chlorpromazine (CPZ), a calmodulin (CAM) inhibitor, drastically suppressed the major catalase isoform as well as total enzymatic activity, and this suppression was alleviated by exogenous sweet potato calmodulin (SPCAM) fusion protein in L3 leaves. CPZ also inhibited the activity of the catalase isoform in vitro. Protein blot hybridization showed that both anti-catalase SPCAT1 and anti-calmodulin SPCAM antibodies detect a band at the same position, which corresponds to the activity of the major catalase isoform from unboiled, but not boiled crude protein extract of L3 leaves. An inverse correlation between the major catalase isoform/total enzymatic activity and the H2O2 level was also observed. These data suggest that sweet potato CAT activity is modulated by CaCl2 and SPCAM, and plays an important role in H2O2 homeostasis in mature leaves. Association of SPCAM with the major CAT isoform is required and regulates the in-gel CAT activity band. PMID:24331417
Afiyanti, Mufidah; Chen, Hsien-Jung
Regulation of catalase activity and gene expression during Phytophthora nicotianae development and infection of tobacco.
Plant defence against pathogen attack typically incorporates an oxidative burst involving elevated levels of reactive oxygen species such as hydrogen peroxide. In the present study, we have used an in-gel assay to monitor the activity of the hydrogen peroxide scavenging enzyme, catalase, during asexual development of Phytophthora nicotianae and during infection of host tobacco plants. In vitro, catalase activity is highest in sporulating hyphae; in planta, catalase activity increases dramatically about 8 h after host inoculation. We have cloned and characterized three catalase genes, designated PnCat1, PnCat2 and PnCat3, from P. nicotianae and identified their homologues in P. infestans, P. sojae and P. ramorum. In all three species, Cat2 is predicted to be targeted to the peroxisome and the other catalases are likely to be cytosolic. Quantitative real-time PCR assessment of catalase transcripts during development and infection indicates that peroxisomal PnCat2 is the gene predominantly expressed, with transcript levels peaking in vitro in sporulating hyphae and in planta increasing dramatically during the first 24 h after inoculation of susceptible tobacco seedlings. Levels of tobacco catalase gene expression are significantly down-regulated in susceptible tobacco 4, 8 and 24 h post-inoculation and in resistant plants at 24 h post-inoculation. Together, our results give evidence that during infection P. nicotianae increases its own peroxisomal catalase levels while concurrently down-regulating host catalase expression. This behaviour is consistent with a role of pathogen catalase in counterdefence and protection against oxidative stress and of pathogen-orchestrated enhanced plant cell death to support necrotrophic pathogen growth and plant colonization. PMID:18705863
Blackman, Leila M; Hardham, Adrienne R
Regulation of cyclo-oxygenase gene expression in rat smooth muscle cells by catalase.
We have studied, in detail, the effect of catalase, one of the naturally occurring antioxidant enzymes, on the expression of cyclo-oxygenase (COX) mRNA and protein in rat aortic smooth muscle cells (RASMC). The activity of COX enzyme within the cells was also determined. Catalase either alone or in combination with interleukin-1beta (IL-1beta) enhanced mRNA and protein expression for cyclo-oxygenase 2 (COX-2) in a concentration-dependent manner. However, it did not affect the expression of mRNA or protein for cyclo-oxygenase 1 (COX-1). The expression of mRNA for COX-2 induced by catalase was blocked completely by actinomycin D (ACT) or cycloheximide (CHX). In comparison, expression of mRNA for COX-2 stimulated by IL-1beta was inhibited by actinomycin D, but not by cycloheximide. This suggests that induction of the synthesis of mRNA for COX-2 by catalase and IL-1beta involves different mechanisms. In particular, the induction of mRNA for COX-2 by catalase requires on-going protein and RNA synthesis, but the induction following exposure to IL-1beta does not. The increase in expression of mRNA for COX-2 induced by catalase may be related to the ability of catalase to stimulate cyclic AMP response element (CRE) and NF-IL6 transcription factors, but not nuclear factor kappa B (NF-kappaB), for electrophoretic mobility shift assays (EMSA) showed that catalase enhanced nuclear factor binding to cyclic AMP response element and NF-IL6 but not to NF-kappaB. Catalase exerted a biphasic effect on prostaglandin synthesis. At low concentrations it enhanced prostaglandin production, but at high concentrations it tended to inhibit it. These findings suggest that catalase has differential and multiple effects on COX expression and activity in rat aortic smooth muscle cells. PMID:9633998
Chen, G; Kamal, M; Hannon, R; Warner, T D
The Monofunctional Catalase KatE of Xanthomonas axonopodis pv. citri Is Required for Full Virulence in Citrus Plants
BackgroundXanthomonas axonopodis pv. citri (Xac) is an obligate aerobic phytopathogen constantly exposed to hydrogen peroxide produced by normal aerobic respiration and by the plant defense response during plant-pathogen interactions. Four putative catalase genes have been identified in silico in the Xac genome, designated as katE, catB, srpA (monofunctional catalases) and katG (bifunctional catalase).Methodology\\/Principal FindingsXac catalase activity was analyzed using native
Mar�a Laura Tondo; Silvana Petrocelli; Jorgelina Ottado; Elena G. Orellano; Ching-Hong Yang
Catalases Promote Resistance of Oxidative Stress in Vibrio cholerae
Oxidative stress is a major challenge faced by bacteria. Many bacteria control oxidative stress resistance pathways through the transcriptional regulator OxyR. The human pathogen Vibrio cholerae is a Gram-negative bacterium that is the causative agent of cholera. V. cholerae lives in both aquatic environments and human small intestines, two environments in which it encounters reactive oxygen species (ROS). To study how V. cholerae responds to oxidative stress, we constructed an in-frame oxyR deletion mutant. We found that this mutant was not only sensitive to H2O2, but also displayed a growth defect when diluted in rich medium. Further study showed that two catalases, KatG and KatB, either when expressed in living cells, present in culture supernatants, or added as purified recombinant proteins, could rescue the oxyR growth defect. Furthermore, although it could colonize infant mouse intestines similar to that of wildtype, the oxyR mutant was defective in zebrafish intestinal colonization. Alternatively, co-infection with wildtype, but not katG-katB deletion mutants, greatly enhanced oxyR mutant colonization. Our study suggests that OxyR in V. cholerae is critical for antioxidant defense and that the organism is capable of scavenging environmental ROS to facilitate population growth.
Rothenbacher, Francesca P.; Jiang, Tiantian; Kan, Biao; Zhong, Zengtao; Zhu, Jun
Enhanced stability of catalase covalently immobilized on functionalized titania submicrospheres.
In this study, a novel approach combing the chelation and covalent binding was explored for facile and efficient enzyme immobilization. The unique capability of titania to chelate with catecholic derivatives at ambient conditions was utilized for titania surface functionalization. The functionalized titania was then used for enzyme immobilization. Titania submicrospheres (500-600 nm) were synthesized by a modified sol-gel method and functionalized with carboxylic acid groups through a facile chelation method by using 3-(3,4-dihydroxyphenyl) propionic acid as the chelating agent. Then, catalase (CAT) was covalently immobilized on these functionalized titania submicrospheres through 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride/N-hydroxysuccinimide (EDC/NHS) coupling reaction. The immobilized CAT retained 65% of its free form activity with a loading capacity of 100-150 mg/g titania. The pH stability, thermostability, recycling stability and storage stability of the immobilized CAT were evaluated. A remarkable enhancement in enzyme stability was achieved. The immobilized CAT retained 90% and 76% of its initial activity after 10 and 16 successive cycles of decomposition of hydrogen peroxide, respectively. Both the Km and the Vmax values of the immobilized CAT (27.4 mM, 13.36 mM/min) were close to those of the free CAT (25.7 mM, 13.46 mM/min). PMID:23827593
Wu, Hong; Liang, Yanpeng; Shi, Jiafu; Wang, Xiaoli; Yang, Dong; Jiang, Zhongyi
A Review on Direct Electrochemistry of Catalase for Electrochemical Sensors
Catalase (CAT) is a heme enzyme with a Fe(III/II) prosthetic group at its redox centre. CAT is present in almost all aerobic living organisms, where it catalyzes the disproportionation of H2O2 into oxygen and water without forming free radicals. In order to study this catalytic mechanism in detail, the direct electrochemistry of CAT has been investigated at various modified electrode surfaces with and without nanomaterials. The results show that CAT immobilized on nanomaterial modified electrodes shows excellent catalytic activity, high sensitivity and the lowest detection limit for H2O2 determination. In the presence of nanomaterials, the direct electron transfer between the heme group of the enzyme and the electrode surface improved significantly. Moreover, the immobilized CAT is highly biocompatible and remains extremely stable within the nanomaterial matrices. This review discusses about the versatile approaches carried out in CAT immobilization for direct electrochemistry and electrochemical sensor development aimed as efficient H2O2 determination. The benefits of immobilizing CAT in nanomaterial matrices have also been highlighted.
Prakash, Periasamy Arun; Yogeswaran, Umasankar; Chen, Shen-Ming
Intron loss and gain during evolution of the catalase gene family in angiosperms.
Angiosperms (flowering plants), including both monocots and dicots, contain small catalase gene families. In the dicot, Arabidopsis thaliana, two catalase (CAT) genes, CAT1 and CAT3, are tightly linked on chromosome 1 and a third, CAT2, which is more similar to CAT1 than to CAT3, is unlinked on chromosome 4. Comparison of positions and numbers of introns among 13 angiosperm catalase genomic sequences indicates that intron positions are conserved, and suggests that an ancestral catalase gene common to monocots and dicots contained seven introns. Arabidopsis CAT2 has seven introns; both CAT1 and CAT3 have six introns in positions conserved with CAT2, but each has lost a different intron. We suggest the following sequence of events during the evolution of the Arabidopsis catalase gene family. An initial duplication of an ancestral catalase gene gave rise to CAT3 and CAT1. CAT1 then served as the template for a second duplication, yielding CAT2. Intron losses from CAT1 and CAT3 followed these duplications. One subclade of monocot catalases has lost all but the 5'-most and 3'-most introns, which is consistent with a mechanism of intron loss by replacement of an ancestral intron-containing gene with a reverse-transcribed DNA copy of a fully spliced mRNA. Following this event of concerted intron loss, the Oryza sativa (rice, a monocot) CAT1 lineage acquired an intron in a novel position, consistent with a mechanism of intron gain at proto-splice sites.
Frugoli, J A; McPeek, M A; Thomas, T L; McClung, C R
Microbial resistance in relation to catalase activity to oxidative stress induced by photolysis of hydrogen peroxide.
The purpose of the present study was to evaluate the mechanism of microbial resistance to oxidative stress induced by photolysis of hydrogen peroxide (H(2)O(2)) in relation to microbial catalase activity. In microbicidal tests, Staphylococcus aureus and Candida albicans were killed and this was accompanied by production of hydroxyl radicals. C. albicans was more resistant to hydroxyl radicals generated by photolysis of H(2)O(2) than was S. aureus. A catalase activity assay demonstrated that C. albicans had stronger catalase activity; accordingly, catalase activity could be one of the reasons for the resistance of the fungus to photolysis of H(2)O(2). Indeed, it was demonstrated that C. albicans with strong catalase activity was more resistant to photolysis of H(2)O(2) than that with weak catalase activity. Kinetic analysis using a modified Lineweaver-Burk plot also demonstrated that the microorganisms reacted directly with hydroxyl radicals and that this was accompanied by decomposition of H(2)O(2). The results of the present study suggest that the microbicidal effects of hydroxyl radicals generated by photolysis of H(2)O(2) can be alleviated by decomposition of H(2)O(2) by catalase in microorganisms. PMID:22040121
Nakamura, Keisuke; Kanno, Taro; Mokudai, Takayuki; Iwasawa, Atsuo; Niwano, Yoshimi; Kohno, Masahiro
Over-Expression of Catalase in Myeloid Cells Confers Acute Protection Following Myocardial Infarction
Cardiovascular disease is the leading cause of death in the United States and new treatment options are greatly needed. Oxidative stress is increased following myocardial infarction and levels of antioxidants decrease, causing imbalance that leads to dysfunction. Therapy involving catalase, the endogenous scavenger of hydrogen peroxide (H2O2), has been met with mixed results. When over-expressed in cardiomyocytes from birth, catalase improves function following injury. When expressed in the same cells in an inducible manner, catalase showed a time-dependent response with no acute benefit, but a chronic benefit due to altered remodeling. In myeloid cells, catalase over-expression reduced angiogenesis during hindlimb ischemia and prevented monocyte migration. In the present study, due to the large inflammatory response following infarction, we examined myeloid-specific catalase over-expression on post-infarct healing. We found a significant increase in catalase levels following infarction that led to a decrease in H2O2 levels, leading to improved acute function. This increase in function could be attributed to reduced infarct size and improved angiogenesis. Despite these initial improvements, there was no improvement in chronic function, likely due to increased fibrosis. These data combined with what has been previously shown underscore the need for temporal, cell-specific catalase delivery as a potential therapeutic option.
Cabigas, E. Bernadette; Somasuntharam, Inthirai; Brown, Milton E.; Che, Pao Lin; Pendergrass, Karl D.; Chiang, Bryce; Taylor, W. Robert; Davis, Michael E.
GM1 ganglioside induces vasodilation and increases catalase content in the brain.
Monosialoganglioside (GM1) is a glycosphingolipid present in most cell membranes that displays antioxidant and neuroprotective properties. GM1 increases catalase activity in cerebral cortices in vivo, but the mechanisms underlying this effect of GM1 are not known. In the current study we investigated the effect of GM1 (50 mg/kg, ip) on the content of hemoglobin and catalase activity of hippocampus, cortex, and striatum of rats. GM1 administration increased catalase activity and hemoglobin content in brain samples after 30 min, but had no effect on blood catalase activity. GM1-induced increase in catalase activity was abolished by brain perfusion with heparinized saline. Brain catalase activity in the absence of blood, estimated by regression analysis of data from perfused and nonperfused animals, was not altered by the systemic injection of GM1. Moreover, the addition of GM1 (30 or 100 microM) did not increase catalase activity in slices of cerebral cortex in situ, further suggesting that blood circulation is required for this effect. The GM1-induced vasodilation was confirmed in vivo, because the systemic injection of GM1 (50 mg/kg, ip) increased (1.2-1.6 times) the width of pial vessels. PMID:17697937
Furian, Ana Fl�via; Oliveira, Mauro Schneider; Royes, Luiz Fernando Freire; Fiorenza, Nat�lia Gindri; Fighera, Michele Rechia; Myskiw, Jociane Carvalho; Weiblen, Rudi; Rubin, Maribel Antonello; Frussa-Filho, Roberto; Mello, Carlos Fernando
Recovery of a Catalase-Negative Staphylococcus epidermidis Strain in Blood and Urine Cultures from a Patient with Pyelonephritis ?
This report describes a 60-year-old patient with bilateral nephrolithiasis. A catalase-negative Staphylococcus epidermidis strain was recovered from both urine and blood cultures. Although rare, isolates of catalase-negative Staphylococcus spp., including Staphylococcus aureus, have been reported. Here, we describe the first report of a catalase-negative S. epidermidis strain.
Kallstrom, George; Chang, Tom; Albertson, Marc; Morilla, Daniel; Fisher, Mark A.; Eberly, Bardwell
Recovery of a catalase-negative Staphylococcus epidermidis strain in blood and urine cultures from a patient with pyelonephritis.
This report describes a 60-year-old patient with bilateral nephrolithiasis. A catalase-negative Staphylococcus epidermidis strain was recovered from both urine and blood cultures. Although rare, isolates of catalase-negative Staphylococcus spp., including Staphylococcus aureus, have been reported. Here, we describe the first report of a catalase-negative S. epidermidis strain. PMID:21900516
Kallstrom, George; Chang, Tom; Albertson, Marc; Morilla, Daniel; Fisher, Mark A; Eberly, Bardwell
Identification of Zantedeschia aethiopica Cat1 and Cat2 catalase genes and their expression analysis during spathe senescence and regreening
Plants encode catalase (CAT; EC 126.96.36.199) as multigene families, which may reflect the multiple and diverse roles played by this enzyme. Catalases from higher plants can be subdivided into three distinct types, according to their phylogenetic relationship. However, there is not a specific correlation of phylogeny and function within these groups, as catalases from the same type can play different
Teresa Lino-Neto; Maria Concei��o Piques; C�tia Barbeta; Manuel F. Sousa; Rui M. Tavares; Maria Salom� Pais
Purification and Characterization of Catalase from Loblolly Pine (Pinus taeda L.) Megagametophytes.
Catalase (EC 220.127.116.11) was purified to near homogeneity from isolated megagametophytes of germinated loblolly pine (Pinus taeda L.) seeds, and monospecific antibodies were elicited in rabbits. Following a procedure that involved acetone extraction, (NH4)2SO4 fractionation, and four chromatographic steps (i.e. DE-52 cellulose, Superdex-200, hydroxylapatite, and phenyl-Sepharose CL-4B), catalase was purified about 140-fold to a final specific activity of 2215 mmol min-1 mg-1 of protein. Cotton isocitrate lyase antibodies were used, and protein immunoblots revealed that the resolution on hydroxylapatite and phenyl-Sepharose allowed for the complete separation of catalase from contaminating isocitrate lyase. The molecular masses of the native enzyme and its subunit are 235 and 59 kD, respectively, indicating that the pine holoenzyme is a homotetramer. Loblolly pine catalase exists as multiple isoforms. When megagametophytes taken 7 d after imbibition at 30[deg]C were extracted, subjected to nondenaturing isoelectric focusing, and stained for catalase activity, at least four catalase isoforms were observed, including one dominant form with an isoelectric point of 6.87. Purified pine catalase is not a glycoprotein and has a ratio of absorbance at 208 nm to absorbance at 405 nm of 1.5. When probed with loblolly pine catalase antibodies, protein blots of cell-free extracts from megagametophytes of mature, stratified, and germinated loblolly pine seeds, the megagametophyte glyoxysomal fraction, and purified loblolly pine catalase all revealed one immunoreactive 59-kD polypeptide. This indicates that no detectable change in the enzyme's monomeric molecular mass occurs during seed stratification and germination, early seedling growth, and purification.
Mullen, R. T.; Gifford, D. J.
Effects of pergolide mesylate on transduction efficiency of PEP-1-catalase protein
Research highlights: {yields} We studied effects of pergolide mesylate (PM) on in vitro and in vivo transduction of PEP-1-catalase. {yields} PEP-1-catatase inhibited 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation. {yields} PM enhanced the transduction of PEP-1-catalase into HaCaT cells and skin tissue. {yields} PM increased anti-inflammatory activity of PEP-1-catalase. {yields} PM stimulated therapeutic action of anti-oxidant enzyme catalase in oxidative-related diseases. -- Abstract: The low transduction efficiency of various proteins is an obstacle to their therapeutic application. However, protein transduction domains (PTDs) are well-known for a highly effective tool for exogenous protein delivery to cells. We examined the effects of pergolide mesylate (PM) on the transduction of PEP-1-catalase into HaCaT human keratinocytes and mice skin and on the anti-inflammatory activity of PEP-1-catatase against 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation using Western blot and histological analysis. PM enhanced the time- and dose-dependent transduction of PEP-1-catalase into HaCaT cells without affecting the cellular toxicity. In a mouse edema model, PEP-1-catalase inhibited the increased expressions of inflammatory mediators and cytokines such as cyclooxygenase-2, inducible nitric oxide synthase, interleukin-6 and -1{beta}, and tumor necrosis factor-{alpha} induced by TPA. On the other hand, PM alone failed to exert any significant anti-inflammatory effects. However, the anti-inflammatory effect of co-treatment with PEP-1-catalase and PM was more potent than that of PEP-1-catalase alone. Our results indicate that PM may enhance the delivery of PTDs fusion therapeutic proteins to target cells and tissues and has potential to increase their therapeutic effects of such drugs against various diseases.
Sohn, Eun Jeong; Kim, Dae Won; Kim, Young Nam; Kim, So Mi [Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702 (Korea, Republic of)] [Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702 (Korea, Republic of); Lim, Soon Sung [Department of Food Science and Nutrition and RIC Center, Hallym University, Chunchon 200-702 (Korea, Republic of)] [Department of Food Science and Nutrition and RIC Center, Hallym University, Chunchon 200-702 (Korea, Republic of); Kang, Tae-Cheon [Department of Anatomy and Neurobiology, College of Medicine, Hallym University, Chunchon 200-702 (Korea, Republic of)] [Department of Anatomy and Neurobiology, College of Medicine, Hallym University, Chunchon 200-702 (Korea, Republic of); Kwon, Hyeok Yil [Department of Physiology, College of Medicine, Hallym University, Chunchon 200-702 (Korea, Republic of)] [Department of Physiology, College of Medicine, Hallym University, Chunchon 200-702 (Korea, Republic of); Kim, Duk-Soo [Department of Anatomy, College of Medicine, Soonchunhyang University, Cheonan-Si 330-090 (Korea, Republic of)] [Department of Anatomy, College of Medicine, Soonchunhyang University, Cheonan-Si 330-090 (Korea, Republic of); Cho, Sung-Woo [Department of Biochemistry and Molecular Biology, University of Ulsan College of Medicine, Seoul 138-736 (Korea, Republic of)] [Department of Biochemistry and Molecular Biology, University of Ulsan College of Medicine, Seoul 138-736 (Korea, Republic of); Han, Kyu Hyung; Park, Jinseu; Eum, Won Sik [Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702 (Korea, Republic of)] [Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702 (Korea, Republic of); Hwang, Hyun Sook, E-mail: firstname.lastname@example.com [Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702 (Korea, Republic of); Choi, Soo Young, E-mail: firstname.lastname@example.com [Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702 (Korea, Republic of)
An ionizable active-site tryptophan imparts catalase activity to a peroxidase core.
Catalase peroxidases (KatG's) are bifunctional heme proteins that can disproportionate hydrogen peroxide (catalatic reaction) despite their structural dissimilarity with monofunctional catalases. Using X-ray crystallography and QM/MM calculations, we demonstrate that the catalatic reaction of KatG's involves deprotonation of the active-site Trp, which plays a role similar to that of the distal His in monofunctional catalases. The interaction of a nearby mobile arginine with the distal Met-Tyr-Trp essential adduct (in/out) acts as an electronic switch, triggering deprotonation of the adduct Trp. PMID:24785434
Loewen, Peter C; Carpena, Xavi; Vidossich, Pietro; Fita, Ignacio; Rovira, Carme
Physicochemical peculiarities of iron porphyrin-containing electrodes in catalase- and peroxidase-type biomimetic sensors
NASA Astrophysics Data System (ADS)
New catalase- and peroxidase-type iron porphyrin biomimetic electrodes have been developed for determining ultralow concentrations of H2O2 and C2H5OH in aqueous solutions. Their physicochemical features have been studied. A mechanism of catalase and peroxidase reactions was suggested. Biomimetic electrodes did not lose their activity for a long time under the action of the oxidant, intermediates, and the final products of the decomposition of H2O2. Potentiometric biomimetic sensors of catalase and peroxidase types have been designed and studied.
Sardarly, N. A.; Nagiev, T. M.
Some kinetic parameters and inactivation of catalase immobilized on modified polyvinylalcohol
Polyvinylalcohol crosslinked with terephthaldicarboxaldehyde and was modified with 2-amino-4,6-dichloro-s-triazine. Optimum\\u000a conditions for immobilization of catalase on modified and gelatine-coated modified polyvinylalcohol were investigated. Activity\\u000a variations with respect to pH, temperature, stability behavior, andk\\u000a m(appl) values were investigated for the native and immobilized catalases. Rate constants for H2O2 decomposition and for inactivation of immobilized catalase were determined using a discontinuous batch-type
Leman Tarhan
Effects of Inflammation on Peroxisomal Enzyme Activity, Catalase Synthesis and Lipid Metabolism.
National Technical Information Service (NTIS)
The activity of hepatic peroxisomal enzymes, catalase, urate oxidase, D-amino acid oxidase, hydroxy acid oxidase, and carnitine acetyltransferase were serially measured following a turpentine-induced sterile inflammatory lesion in rats. The ensuing depres...
P. G. Canonico W. Rill E. Ayala
Studies of Peroxide Deactivation and Glucose Removal with Immobilized Glucose Oxidase and Catalase.
National Technical Information Service (NTIS)
A series of studies using immobilized glucose oxidase and catalase in a continuous flow, tubular, packed bed reactor have been performed. The studies were aimed at developing an immobilized enzyme system for the desugaring of eggs. Quantitative expression...
R. E. Altomare
A Simple Method for Demonstrating Enzyme Kinetics Using Catalase from Beef Liver Extract
NASA Astrophysics Data System (ADS)
This paper describes a simple visual method of demonstrating enzyme kinetics using beef liver catalase. A catalase solution is obtained by homogenizing beef liver in a phosphate buffer. In the demonstration, filter paper is saturated with beef liver extract and placed into a solution of hydrogen peroxide. The catalase in the extract decomposes the hydrogen peroxide to water and oxygen. Oxygen forms on the filter paper, and the filter paper rises to the top of the beaker. Catalase activity is measured by timing the rise of the enzyme-soaked filter paper to the top of beakers containing different concentrations of hydrogen peroxide. The data are plotted as a Lineweaver-Burk double-reciprocal plot, and the Km and Vmax for the reaction are calculated.
Johnson, Kristin A.
Serum adenosine deaminase, catalase and carbonic anhydrase activities in patients with bladder cancer
OBJECTIVES: The relationship between adenosine deaminase and various cancers has been investigated in several studies. However, serum adenosine deaminase activity and carbonic anhydrase and catalase activities in patients with bladder cancer have not previously been reported. Therefore, the aim of this study was to measure serum adenosine deaminase, carbonic anhydrase and catalase activities in patients with bladder cancer. MATERIALS AND METHODS: Forty patients with bladder cancer and 30 healthy controls were enrolled in the study. Serum adenosine deaminase, carbonic anhydrase and catalase activities were measured spectrophotometrically. RESULTS: Serum adenosine deaminase, carbonic anhydrase and catalase activities were significantly higher in patients with bladder cancer than controls (all significant, p<0.001). CONCLUSIONS: These markers might be a potentially important finding as an additional diagnostic biochemical tool for bladder cancer.
Pirincci, Necip; Gecit, Ilhan; Gunes, Mustafa; Bilgehan Y?ksel, Mehmet; Kaba, Mehmet; Tan?k, Serhat; Demir, Halit; Aslan, Mehmet
[Catalytic properties and stability of catalase in reversed micelles of aerosol OT in octane].
The catalytic function of catalase and its peroxidatic activity during tetramethylbenzidine (TMB) oxidation by cumene hydroperoxide were studied in reversed micelles of Aerosol OT (AOT) in octane relative to the [H2O]/[AOT] ratio and the initial catalase concentration. The optimum conditions permitting to retain the catalytic activity of the enzyme and its ability to induce peroxidation of TMB, were found. The catalytic function of the enzyme was shown to be dependent on its concentration in AOT micelles. The catalase stability monitored by the catalytic reaction and the decrease of the Soret band were analyzed. Both processes have two phases differing by the rate constants of the pseudo-first order. The catalase inserted into AOT micelles is characterized by the high stability as compared to other hemoproteins (cytochrome P-450, myoglobin, hemoglobin, peroxidase) under identical conditions. PMID:1692742
Artemchik, V D; Eremin, A N; Metelitsa, D I
[Stability of Penicillium vitale immobilized catalase in continuous decomposition of hydrogen peroxide].
The process of hydrogen peroxide continuous decomposition by the preparation of the fungus Penicillium vitale catalase immobilized by aminoorganosilica which were activated by glutaric aldehyde, cyanuric chloride or 2,4-toluylene diisocyanate. Catalase with an oxidized carbohydrate component was used as well. Such a modified enzyme was directly bound with the surface of aminocontaining silica and alumina. It is shown that in the process of H2O2 decomposition the preparations of immobilized catalase are inactivated. The decrease in its activity is described by a model which suggests that rates of hydrogen peroxide decomposition and enzyme inactivation are described by the first order equations. A method for calculation and prediction of mean time of continuous operation of columns with bound catalase and other immobilized enzymes is suggested in terms of the given model. PMID:7281255
Ianishpol'ski?, V V; Tertykh, V A; Gudkova, L V; Latyshko, N V
Catalase-dependent peroxidative metabolism in the alveolar macrophage during phagocytosis
Evidence for the presence of peroxidative metabolism in rabbit alveolar macrophages (AM) has been obtained from the following observations: (a) catalase is present in high concentrations; (b) peroxidase activity could not be detected employing guaiacol as substrate; (c) the irreversible inhibition of AM catalase by aminotriazole served as a detection system for H2O2 and demonstrated increased intracellular H2O2 after phagocytosis; (d) formate oxidation, a marker of catalase-dependent peroxidations, occurs in resting AM and is increased by phagocytosis; (c) measurements of H2O2 accumulation in a dialysate of AM demonstrated twofold increase during phagocytosis; and (f) aminotriazole diminishes O2 utilization and 14CO2 production from labelled glucose and pyruvate. It is concluded that, while catalase-dependent H2O2 metabolism is not essential for particle entry, this pathway represents one of the metabolic pathways stimulated by particle entry in the AM.
Gee, J. Bernard L.; Vassallo, Charles L.; Bell, Paul; Kaskin, James; Basford, R. E.; Field, James B.
Production, characterization, cloning and sequence analysis of a monofunctional catalase from Serratia marcescens SYBC08.
A monofunctional catalase from Serratia marcescens SYBC08 produced by liquid state fermentation in 7 liter fermenter was isolated and purified by ammonium sulfate precipitation (ASP), ion exchange chromatography (IEC), and gel filtration (GF) and characterized. Its sequence was analyzed by LC-MS/MS technique and gene cloning. The highest catalase production (20,289 U � ml(-1)) was achieved after incubation for 40 h. The purified catalase had an estimated molecular mass of 230 kDa, consisting of four identical subunits of 58 kDa. High specific activity of the catalase (199,584 U � mg(-1) protein) was 3.44 times higher than that of Halomonas sp. Sk1 catalase (57,900 U � mg(-1) protein). The enzyme without peroxidase activity was found to be an atypical electronic spectrum of monofunctional catalase. The apparent K(m) and V(max) were 78 mM and 188, 212 per �M H(2) O(2) �M heme(-1) s(-1), respectivly. The enzyme displayed a broad pH activity range (pH 5.0-11.0), with optimal pH range of 7.0-9.0: It was most active at 20 �C and had 78% activity at 0 �C. Its thermo stability was slightly higher compared to that of commercial catalase from bovine liver. LC-MS/MS analysis confirmed that the deduced amino acid sequence of cloning gene was the catalase sequence from Serratia marcescens SYBC08. The sequence was compared with that of 23 related catalases. Although most of active site residues, NADPH-binding residues, proximal residues of the heme, distal residues of the heme and residues interacting with a water molecule in the enzyme were well conserved in 23 related catalases, weakly conserved residues were found. Its sequence was closely related with that of catalases from pathogenic bacterium in the family Enterobacteriaceae. This result imply that the enzyme with high specific activity plays a significant role in preventing those microorganisms of the family Enterobacteriaceae against hydrogen peroxide resulted in cellular damage. Calalase yield by Serratia marcescens SYBC08 has potential industrial application in scavenging hydrogen peroxide. PMID:21077118
Zeng, Hua-Wei; Cai, Yu-Jie; Liao, Xiang-Ru; Zhang, Feng; Zhang, Da-Bing
The circadian clock gates expression of two Arabidopsis catalase genes to distinct and opposite circadian phases
InArabidopsis thaliana, catalase is encoded by a small gene family. We have characterized cDNA and genomic clones containing theArabidopsis catalase geneCAT3, present as a single copy in the nuclear genome. Six introns were identified in theCAT3 coding region and two transcription start sites have been been mapped by primer extension. The deduced amino acid sequence of CAT3 is highly similar
H. H. Zhong; C. R. McClung
Purification, characterization, and identification of a novel bifunctional catalase-phenol oxidase from Scytalidium thermophilum
A novel bifunctional catalase with an additional phenol oxidase activity was isolated from a thermophilic fungus, Scytalidium thermophilum. This extracellular enzyme was purified ca. 10-fold with 46% yield and was biochemically characterized. The enzyme contains\\u000a heme and has a molecular weight of 320�kDa with four 80�kDa subunits and an isoelectric point of 5.0. Catalase and phenol\\u000a oxidase activities were most
Didem Sutay Kocabas; Ufuk Bakir; Simon E. V. Phillips; Michael J. McPherson; Zumrut B. Ogel
Catalase-negative Actinomyces neuii subsp. neuii isolated from an infected mammary prosthesis
In this case report a catalase-negative strain of Actinomyces neuii subsp. neuii is described as the possible causative agent of an infected mammary prosthesis. DNA hybridization studies and 16S rRNA analysis confirmed that the strain belongs to the species Actinomyces neuii subsp. neuii. Since this strain is the first A. neuii subsp. neuii strain reported to be catalase negative, the
Simone Brunner; Susanne Graf; Philippe Riegel; Martin Altwegg
Physicochemical peculiarities of iron porphyrin-containing electrodes in catalase- and peroxidase-type biomimetic sensors
New catalase- and peroxidase-type iron porphyrin biomimetic electrodes have been developed for determining ultralow concentrations of H2O2 and C2H5OH in aqueous solutions. Their physicochemical features have been studied. A mechanism of catalase and peroxidase reactions was suggested. Biomimetic electrodes did not lose their activity for a long time under the action of the oxidant, intermediates, and the final products of
N. A. Sardarly; T. M. Nagiev
Physicochemical peculiarities of iron porphyrin-containing electrodes in catalase- and peroxidase-type biomimetic sensors
New catalase- and peroxidase-type iron porphyrin biomimetic electrodes have been developed for determining ultralow concentrations\\u000a of H2O2 and C2H5OH in aqueous solutions. Their physicochemical features have been studied. A mechanism of catalase and peroxidase reactions\\u000a was suggested. Biomimetic electrodes did not lose their activity for a long time under the action of the oxidant, intermediates,\\u000a and the final products of
N. A. Sardarly; T. M. Nagiev
Separation of glucose oxidase and catalase using ultrafiltration with 300-kDa polyethersulfone membranes
This work examines the separation of glucose oxidase (GOx) and catalase by ultrafiltration using 300kDa MWCO polyethersulfone membranes. The effects of solution pH and ionic strength on the transmission of a single protein (either GOx or catalase) were quantified using the pulsed sample injection technique. Other effects including stirring speed and permeate flux on separation of the binary protein mixtures
Jianguo Liu; Junren Lu; Xiubo Zhao; Jianren Lu; Zhanfeng Cui
Layer-by-layer assembled multilayers using catalase-encapsulated gold nanoparticles
We introduce a novel and versatile approach for the preparation of multilayers, based on catalase-encapsulated gold nanoparticles (CAT-AuNP), allowing electrostatic charge reversal and structural transformation through pH adjustment. CAT-AuNP, which are synthesized directly from CAT stabilizer, can be electrostatically assembled with anionic and cationic PEs as a result of the charge reversal of the catalase stabilizers through pH control. In
Sungwoo Kim; Jinhan Cho
Regulation of Cyclo-oxygenase Gene Expression in Rat Smooth Muscle Cells by Catalase
We have studied, in detail, the effect of catalase, one of the naturally occurring antioxidant enzymes, on the expression of cyclo-oxygenase (COX) mRNA and protein in rat aortic smooth muscle cells (RASMC). The activity of COX enzyme within the cells was also determined. Catalase either alone or in combination with interleukin-1? (IL-1?) enhanced mRNA and protein expression for cyclo-oxygenase 2
Gong Chen; Mahine Kamal; Robert Hannon; Timothy D. Warner
Separation and characterization of two catalase activities isolated from the yeast Trigonopsis variabilis
Two different catalases present in cell-free extracts of the yeast Trigonopsis variabilis have been separated by dye-binding chromatography. Both enzymes, named TvC-I and TvC-II, behave as members of the �typical catalases� family: they have a broad pH activity profile and do not possess peroxidase activity, are quite stable when treated with ethanol\\/chloroform mixtures, are inhibited by azide and cyanide at
Daniela Monti; Eva Baldaro; Sergio Riva
Increased expression of catalase and superoxide dismutase 2 reduces cone cell death in retinitis pigmentosa.
Oxidative and nitrosative damage are major contributors to cone cell death in retinitis pigmentosa (RP). In this study, we explored the effects of augmenting components of the endogenous antioxidant defense system in models of RP, rd1, and rd10 mice. Unexpectedly, overexpression of superoxide dismutase 1 (SOD1) in rd1 mice increased oxidative damage and accelerated cone cell death. With an elaborate mating scheme, genetically engineered rd10 mice with either inducible expression of SOD2, Catalase, or both in photoreceptor mitochondria were generated. Littermates with the same genetic background that did not have increased expression of SOD2 nor Catalase provided ideal controls. Coexpression of SOD2 and Catalase, but not either alone, significantly reduced oxidative damage in the retinas of postnatal day (P) 50 rd10 mice as measured by protein carbonyl content. Cone density was significantly greater in P50 rd10 mice with coexpression of SOD2 and Catalase together than rd10 mice that expressed SOD2 or Catalase alone, or expressed neither. Coexpression of SOD2 and Catalase in rd10 mice did not slow rod cell death. These data support the concept of bolstering the endogenous antioxidant defense system as a gene-based treatment strategy for RP, and also indicate that coexpression of multiple components may be needed. PMID:19293779
Usui, Shinichi; Komeima, Keiichi; Lee, Sun Young; Jo, Young-Joon; Ueno, Shinji; Rogers, Brian S; Wu, Zhihao; Shen, Jikui; Lu, Lili; Oveson, Brian C; Rabinovitch, Peter S; Campochiaro, Peter A
The critical role of catalase in prooxidant and antioxidant function of p53
The tumor suppressor p53 is an important regulator of intracellular reactive oxygen species (ROS) levels, although downstream mediators of p53 remain to be elucidated. Here, we show that p53 and its downstream targets, p53-inducible ribonucleotide reductase (p53R2) and p53-inducible gene 3 (PIG3), physically and functionally interact with catalase for efficient regulation of intracellular ROS, depending on stress intensity. Under physiological conditions, the antioxidant functions of p53 are mediated by p53R2, which maintains increased catalase activity and thereby protects against endogenous ROS. After genotoxic stress, high levels of p53 and PIG3 cooperate to inhibit catalase activity, leading to a shift in the oxidant/antioxidant balance toward an oxidative status, which could augment apoptotic cell death. These results highlight the essential role of catalase in p53-mediated ROS regulation and suggest that the p53/p53R2�catalase and p53/PIG3�catalase pathways are critically involved in intracellular ROS regulation under physiological conditions and during the response to DNA damage, respectively.
Kang, M Y; Kim, H-B; Piao, C; Lee, K H; Hyun, J W; Chang, I-Y; You, H J
Leptospira interrogans Catalase Is Required for Resistance to H2O2 and for Virulence
Pathogenic Leptospira spp. are likely to encounter higher concentrations of reactive oxygen species induced by the host innate immune response. In this study, we characterized Leptospira interrogans catalase (KatE), the only annotated catalase found within pathogenic Leptospira species, by assessing its role in resistance to H2O2-induced oxidative stress and during infection in hamsters. Pathogenic L. interrogans bacteria had a 50-fold-higher survival rate under H2O2-induced oxidative stress than did saprophytic L. biflexa bacteria, and this was predominantly catalase dependent. We also characterized KatE, the only annotated catalase found within pathogenic Leptospira species. Catalase assays performed with recombinant KatE confirmed specific catalase activity, while protein fractionation experiments localized KatE to the bacterial periplasmic space. The insertional inactivation of katE in pathogenic Leptospira bacteria drastically diminished leptospiral viability in the presence of extracellular H2O2 and reduced virulence in an acute-infection model. Combined, these results suggest that L. interrogans KatE confers in vivo resistance to reactive oxygen species induced by the host innate immune response.
Eshghi, Azad; Lourdault, Kristel; Murray, Gerald L.; Bartpho, Thanatchaporn; Sermswan, Rasana W.; Picardeau, Mathieu; Adler, Ben; Snarr, Brendan; Zuerner, Richard L.
Determination of catalase activity using chromogenic probe involving iso-nicotinicacidhydrazide and pyrocatechol.
A biocatalatic pathway involving chromogenic probe has been proposed for the determination of catalase activity by means of iso-nicotinicacidhydrazide (INH) and pyrocatechol (PC). The assay is based on the enzymatic consumption of hydrogen peroxide using INH-PC system. The response of the catalase activity was ascertained by the rate of the reaction involving 14.10mM H(2)O(2). On addition of H(2)O(2), INH-PC indicator system formed a chromogenic product with absorbance maxima at 490 nm. Hence the activity of catalase was directly measured by the chromogenic response in the formation of the coupled product. The catalase assay was elaborated by the kinetic response of the INH-PC system. The linearity of the catalase activity and H(2)O(2) was in the range 0.2-7.0 units and 1.76-7.0mM, respectively in 3 ml solution. The catalytic efficiency and catalytic power were calculated. The Michaelis-Menten constant of INH, PC and H(2)O(2) were found to be 0.344, 0.176 and 8.82 mM, respectively. The indicator reaction was applied in the determination of catalase activity in mycelia mats and culture media. PMID:21839122
Shivakumar, Anantharaman; Nagaraja, Padmarajaiah; Chamaraja, Nelligere Arkeshwaraiah; Krishna, Honnur; Avinash, Krishnegowda
Production and some properties of catalase and superoxide dismutase from the anaerobe Bacteroides distasonis.
The catalase level of Bacteroides distasonis (ATCC 8503, type strain) varied with the amount of hemin supplied to the medium when the cells were grown in either a prereduced medium containing 0.5% peptone, 0.5% yeast extract, and 1% glucose or in a prereduced, defined heme-deficient medium. The effect of hemin on catalase production could not be duplicated by ferrous sulfate or ferrous ammonium citrate. Catalase activity reached peak values in late log phase, whereas superoxide dismutase specific activity remained constant throughout the culture growth cycle. The catalase was a nondialyzable, cyanide and azide-sensitive, heat-labile protein that coeluted with bovine erythrocyte catalase from Sepharose 6 B. Analysis of polyacrylamide gels stained for catalase activity and for heme showed a correspondence between the single catalytic activity band and one of three heme-protein bands. These data suggest a heme-protein of approximately 250,000 molecular weight. The superoxide dismutase was a cyanide-insensitive protein of approximately 40,000 molecular weight that migrated electrophoretically on acrylamide gels as a single band of activity.
Gregory, E M; Kowalski, J B; Holdeman, L V
Nitrite-catalase interaction as an important element of nitrite toxicity.
It was established that nitrite in the presence of chloride, bromide, and thiocyanate decreases the rate of hydrogen peroxide decomposition by catalase. The decrease was recorded by the permanganatometric method and by a method of dynamic calorimetry. Nitrite was not destroyed in the course of the reaction and the total value of heat produced in the process was not changed by its presence. These facts suggest that nitrite induces inhibition of catalase with no change in the essence of the enzymatic process. Even micromolar nitrite concentrations induced a considerable decrease in catalase activity. However, in the absence of chloride, bromide, and thiocyanate inhibition was not observed. In contrast, fluoride protected catalase from nitrite inhibition in the presence of the above-mentioned halides and pseudohalide. As hydrogen peroxide is a necessary factor for triggering a number of important toxic effects of nitrite, the latter increases its toxicity by inhibiting catalase. This was shown by the example of nitrite-induced hemoglobin oxidation. The naturally existing gradient of chloride and other anion concentrations between intra- and extracellular media appears to be the most important mechanism of cell protection from inhibition of intracellular catalase by nitrite. Possible mechanisms of this inhibition are discussed. PMID:12943506
Titov, V Yu; Petrenko, Yu M
Redundant catalases detoxify phagocyte reactive oxygen and facilitate Histoplasma capsulatum pathogenesis.
Histoplasma capsulatum is a respiratory pathogen that infects phagocytic cells. The mechanisms allowing Histoplasma to overcome toxic reactive oxygen molecules produced by the innate immune system are an integral part of Histoplasma's ability to survive during infection. To probe the contribution of Histoplasma catalases in oxidative stress defense, we created and analyzed the virulence defects of mutants lacking CatB and CatP, which are responsible for extracellular and intracellular catalase activities, respectively. Both CatB and CatP protected Histoplasma from peroxide challenge in vitro and from antimicrobial reactive oxygen produced by human neutrophils and activated macrophages. Optimal protection required both catalases, as the survival of a double mutant lacking both CatB and CatP was lower than that of single-catalase-deficient cells. Although CatB contributed to reactive oxygen species defenses in vitro, CatB was dispensable for lung infection and extrapulmonary dissemination in vivo. Loss of CatB from a strain also lacking superoxide dismutase (Sod3) did not further reduce the survival of Histoplasma yeasts. Nevertheless, some catalase function was required for pathogenesis since simultaneous loss of both CatB and CatP attenuated Histoplasma virulence in vivo. These results demonstrate that Histoplasma's dual catalases comprise a system that enables Histoplasma to efficiently overcome the reactive oxygen produced by the innate immune system. PMID:23589579
Holbrook, Eric D; Smolnycki, Katherine A; Youseff, Brian H; Rappleye, Chad A
Association of catalase gene polymorphisms with catalase activity and susceptibility to systemic lupus erythematosus in the Suez Canal area, Egypt.
The present study evaluated the relationship of genetic variants in both promoter (-262?C/T) and in exonic (389?C/T) regions of the catalase (CAT) gene to CAT activity and risk of systemic lupus erythematosus (SLE) in Suez Canal-area patients. CAT gene polymorphisms were assessed by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). CAT activity was measured by using a spectrophotometer. We compared the frequencies of CAT 389 C/T and -262?C/T polymorphic variants between SLE patients (n?=?103) and healthy controls (n?=?103). CAT 389?C/T is associated with SLE susceptibility, with the T allele being significantly more frequent among SLE patients than healthy controls. There was no association, however, between CAT activity and genotypes of 389?C/T. We did not observe significant differences in the prevalence of CAT -262?C/T polymorphic variants in SLE patients and controls, however, we found that patients with the CAT -262 CT and TT genotypes had low CAT activity, and these genotypes showed a significant association with thrombocytopaenia, leukopaenia and the presence of anti-snRNP in SLE patients. In conclusion, the present study supports the notion of in�vivo oxidative stress in SLE as indicated by the decrease in CAT activity. The allelic variations in the CAT gene -262 are more likely to affect the expression or the function of the enzyme. Since CAT may be pathogenetically linked to SLE, and owing to its free-radical origin, it appears reasonable to target lipid peroxidation by dietary and/or pharmacological antioxidants. PMID:22736749
Ghaly, M S; Ghattas, M H; Labib, S M
Double antisense plants lacking ascorbate peroxidase and catalase are less sensitive to oxidative stress than single antisense plants lacking ascorbate peroxidase or catalase
Summary The plant genome is a highly redundant and dynamic genome. Here, we show that double antisense plants lacking the two major hydrogen peroxide-detoxifying enzymes, ascorbate peroxidase (APX) and catalase (CAT), activate an alternative\\/redundant defense mechanism that compensates for the lack of APX and CAT. A similar mechanism was not activated in single antisense plants that lacked APX or CAT,
Ludmila Rizhsky; Elza Hallak-Herr; Frank Van Breusegem; Shimon Rachmilevitch; Jason E. Barr; Steven Rodermel; Dirk Inz�; Ron Mittler
Catalase is a sink for H2O2 and is indispensable for stress defence in C3 plants.
Hydrogen peroxide (H2O2) has been implicated in many stress conditions. Control of H2O2 levels is complex and dissection of mechanisms generating and relieving H2O2 stress is difficult, particularly in intact plants. We have used transgenic tobacco with approximately 10% wild-type catalase activity to study the role of catalase and effects of H2O2 stress in plants. Catalase-deficient plants showed no visible disorders at low light, but in elevated light rapidly developed white necrotic lesions on the leaves. Lesion formation required photorespiratory activity since damage was prevented under elevated CO2. Accumulation of H2O2 was not detected during leaf necrosis. Alternative H2O2-scavenging mechanisms may have compensated for reduced catalase activity, as shown by increased ascorbate peroxidase and glutathione peroxidase levels. Leaf necrosis correlated with accumulation of oxidized glutathione and a 4-fold decrease in ascorbate, indicating that catalase is critical for maintaining the redox balance during oxidative stress. Such control may not be limited to peroxisomal H2O2 production. Catalase functions as a cellular sink for H2O2, as evidenced by complementation of catalase deficiency by exogenous catalase, and comparison of catalase-deficient and control leaf discs in removing external H2O2. Stress analysis revealed increased susceptibility of catalase-deficient plants to paraquat, salt and ozone, but not to chilling. PMID:9305623
Willekens, H; Chamnongpol, S; Davey, M; Schraudner, M; Langebartels, C; Van Montagu, M; Inz�, D; Van Camp, W
Catalase is a sink for H2O2 and is indispensable for stress defence in C3 plants.
Hydrogen peroxide (H2O2) has been implicated in many stress conditions. Control of H2O2 levels is complex and dissection of mechanisms generating and relieving H2O2 stress is difficult, particularly in intact plants. We have used transgenic tobacco with approximately 10% wild-type catalase activity to study the role of catalase and effects of H2O2 stress in plants. Catalase-deficient plants showed no visible disorders at low light, but in elevated light rapidly developed white necrotic lesions on the leaves. Lesion formation required photorespiratory activity since damage was prevented under elevated CO2. Accumulation of H2O2 was not detected during leaf necrosis. Alternative H2O2-scavenging mechanisms may have compensated for reduced catalase activity, as shown by increased ascorbate peroxidase and glutathione peroxidase levels. Leaf necrosis correlated with accumulation of oxidized glutathione and a 4-fold decrease in ascorbate, indicating that catalase is critical for maintaining the redox balance during oxidative stress. Such control may not be limited to peroxisomal H2O2 production. Catalase functions as a cellular sink for H2O2, as evidenced by complementation of catalase deficiency by exogenous catalase, and comparison of catalase-deficient and control leaf discs in removing external H2O2. Stress analysis revealed increased susceptibility of catalase-deficient plants to paraquat, salt and ozone, but not to chilling.
Willekens, H; Chamnongpol, S; Davey, M; Schraudner, M; Langebartels, C; Van Montagu, M; Inze, D; Van Camp, W
Adeno-Associated Viral-Mediated Catalase Expression Suppresses Optic Neuritis in Experimental Allergic Encephalomyelitis
NASA Astrophysics Data System (ADS)
Suppression of oxidative injury by viral-mediated transfer of the human catalase gene was tested in the optic nerves of animals with experimental allergic encephalomyelitis (EAE). EAE is an inflammatory autoimmune disorder of primary central nervous system demyelination that has been frequently used as an animal model for the human disease multiple sclerosis (MS). The optic nerve is a frequent site of involvement common to both EAE and MS. Recombinant adeno-associated virus containing the human gene for catalase was injected over the right optic nerve heads of SJL/J mice that were simultaneously sensitized for EAE. After 1 month, cell-specific catalase activity, evaluated by quantitation of catalase immunogold, was increased approximately 2-fold each in endothelia, oligodendroglia, astrocytes, and axons of the optic nerve. Effects of catalase on the histologic lesions of EAE were measured by computerized analysis of the myelin sheath area (for demyelination), optic disc area (for optic nerve head swelling), extent of the cellular infiltrate, extravasated serum albumin labeled by immunogold (for blood-brain barrier disruption), and in vivo H2O2 reaction product. Relative to control, contralateral optic nerves injected with the recombinant virus without a therapeutic gene, catalase gene inoculation reduced demyelination by 38%, optic nerve head swelling by 29%, cellular infiltration by 34%, disruption of the blood-brain barrier by 64%, and in vivo levels of H2O2 by 61%. Because the efficacy of potential treatments for MS are usually initially tested in the EAE animal model, this study suggests that catalase gene delivery by using viral vectors may be a therapeutic strategy for suppression of MS.
Guy, John; Qi, Xiaoping; Hauswirth, William W.
Restoring catalase activity in Staphylococcus aureus subsp. anaerobius leads to loss of pathogenicity for lambs.
Staphylococcus aureus subsp. anaerobius, a microaerophilic and catalase-negative bacterium, is the etiological agent of abscess disease, a specific chronic condition of sheep and goats, which is characterized by formation of necrotic lesions that are located typically in superficial lymph nodes. We constructed an isogenic mutant of S. aureus subsp. anaerobius (RDKA84) that carried a repaired and functional catalase gene from S. aureus ATCC 12600, to investigate whether the lack of catalase in S. aureus subsp. anaerobius plays a role in its physiological and pathogenic characteristics. The catalase activity had no apparent influence on the in vitro growth characteristics of RDKA84, which, like the wild-type, did not grow on aerobically incubated agar plates. Restoration of catalase activity in RDKA84 substantially increased resistance to H2O2 when analyzed in a death assay. The intracellular survival rates of the catalase-positive mutant RDKA84 in polymorphonuclear neutrophils (PMN) isolated from adult sheep were significantly higher than those of the wild-type, while no differences were found with PMN isolated from lambs. RDKA84 showed significantly lower survival rates in murine macrophages (J774A.1 cells) than the wild-type strains did, whereas, in bovine mammary epithelial cells (MAC-T), no differences in intracellular survival were observed. Interestingly, the virulence for lambs, the natural host for abscess disease, of the catalase-positive mutant RDKA84 was reduced dramatically in comparison with wild-type S. aureus subsp. anaerobius in two experimental models of infection. PMID:20167202
de la Fuente, Ricardo; D�ez, Rosa M; Dom�nguez-Bernal, Gustavo; Orden, Jos� A; Mart�nez-Pulgar�n, Susana
Protection of Bacillus larvae from Oxygen Toxicity with Emphasis on the Role of Catalase
Sporulation of Bacillus larvae NRRL B-3650 occurred only at aeration rates lower than those used for cultivation of most Bacillus species. One possible explanation for the requirement for a low level of aeration in B. larvae is that toxic forms of oxygen such as H2O2 and superoxide are involved. The superoxide dismutase levels of strain B-3650 were similar to those of Bacillus subtilis 168 during sporulation, and no NADH peroxidase was present. Catalase activity was absent during exponential growth and first appeared near the start of the stationary phase. The catalase activity was 2,700 times less than that in B. subtilis 168 at the same stage of development. Therefore, the relative deficiency of catalase (and NADH peroxidase) might be the cause of the apparent O2 toxicity. It was postulated that B. larvae might accumulate H2O2 in the medium and exhibit more than normal sensitivity to H2O2. Experimental results did not verify either postulate, but the possibilities of intracellular accumulation of H2O2 and unusual sensitivity to endogenous H2O2 were not excluded. The catalase present in early-stationary-phase cells was soluble, heat labile, and inhibited by cyanide, azide, and hydroxylamine. An increase in catalase activity also occurred at the time of appearance of refractile spores in both B. larvae NRRL B-3650 and B. subtilis 168. The level of catalase activity in strain B-3650 was 5,400 times less than that in B. subtilis 168 at this stage. In B. larvae, this second increase occurred primarily within the developing endospore. The activity in spore extracts was particulate, heat stable, and inhibited by hydroxylamine but not by azide or cyanide. Synthesis of catalase in B. larvae was unaffected by H2O2, O2, or glucose.
Dingman, D. W.; Stahly, Donald P.
Development of a new catalase activity assay for biological samples using optical CUPRAC sensor
NASA Astrophysics Data System (ADS)
A novel catalase activity assay was developed for biological samples (liver and kidney tissue homogenates) using a rapid and low-cost optical sensor-based �cupric reducing antioxidant capacity� (CUPRAC) method. The reagent, copper(II)-neocuproine (Cu(II)-Nc) complex, was immobilized onto a cation-exchanger film of Nafion, and the absorbance changes associated with the formation of the highly-colored Cu(I)-Nc chelate as a result of reaction with hydrogen peroxide (H2O2) was measured at 450 nm. When catalase was absent, H2O2 produced the CUPRAC chromophore, whereas catalase, being an effective H2O2 scavenger, completely annihilated the CUPRAC signal due to H2O2. Thus, the CUPRAC absorbance due to H2O2 oxidation concomitant with Cu(I)-Nc formation decreased proportionally with catalase. The developed sensor gave a linear response over a wide concentration range of H2O2 (0.68-78.6 ?M). This optical sensor-based method applicable to tissue homogenates proved to be efficient for low hydrogen peroxide concentrations (physiological and nontoxic levels) to which the widely used UV method is not accurately responsive. Thus, conventional problems of the UV method arising from relatively low sensitivity and selectivity, and absorbance disturbance due to gaseous oxygen evolution were overcome. The catalase findings of the proposed method for tissue homogenates were statistically alike with those of HPLC.
Bekde?er, Burcu; �zy�rek, Mustafa; G��l�, Kubilay; Alkan, Fulya �st�n; Apak, Re?at
The oxidation of chiral alcohols catalyzed by catalase in organic solvents
The catalytic properties of bovine liver catalase have been investigated in organic solvents. In tetrahydrofuran, dioxane, and acetone (all containing 1% to 3% of water), the enzyme breaks down tert-butyl hydroperoxide several fold faster than in pure water. Furthermore, the rate of catalase-catalyzed production of tert-butanol from tert-butyl hydroperoxide increases more than 400-fold upon transition from aqueous buffer to ethanol as the reaction medium. The mechanistic rationale for this striking effect is that in aqueous buffer the rate-limiting step of the enzymatic process involves the reduction of catalase`s compound 1 by tert-butyl hydroperoxide. In ethanol, an additional step in the reaction scheme becomes available in which ethanol, greatly outcompeting the hydroperoxide, is oxidized by compound 1 regenerating the free enzyme. In solvents, such as acetonitrile or tetrahydrofuran, which themselves are not oxidizable by compound 1, catalase catalyzes the oxidation of numerous primary and secondary alcohols with tert-butyl hydroperoxide to the corresponding aldehydes or ketones. The enzymatic oxidation of some chiral alcohols (2,3-butanediol, citronellol, and menthol) under these conditions occurs enantioselectively. Examination of the enantioselectivity for the oxidation of 2,3-butanediol in a series of organic solvents reveals a considerable solvent dependence.
Magner, E.; Klibanov, A.M. [Massachusetts Inst. of Tech., Cambridge, MA (United States). Dept. of Chemistry
Immobilization and kinetics of catalase on calcium carbonate nanoparticles attached epoxy support.
A novel hybrid epoxy/nano CaCO3 composite matrix for catalase immobilization was prepared by polymerizing epoxy resin in the presence of CaCO3 nanoparticles. The hybrid support was characterized using scanning electron microscopy and Fourier transform infrared spectroscopy. Catalase was successfully immobilized onto epoxy/nano CaCO3 support with a conjugation yield of 0.67 � 0.01 mg/cm(2) and 92.63 � 0.80 % retention of activity. Optimum pH and optimum temperature of free and immobilized catalases were found to be 7.0 and 35 �C. The value of Km for H2O2 was higher for immobilized enzyme (31.42 mM) than native enzyme (27.73 mM). A decrease in Vmax value from 1,500 to 421.10 ?mol (min mg protein)(-1) was observed after immobilization. Thermal and storage stabilities of catalase improved immensely after immobilization. Immobilized enzyme retained three times than the activity of free enzyme when kept at 75 �C for 1 h and the half-life of enzyme increased five times when stored in phosphate buffer (0.01 M, pH 7.0) at 5 �C. The enzyme could be reused 30 times without any significant loss of its initial activity. Desorption of catalase from the hybrid support was minimum at pH 7.0. PMID:24048961
Preety; Hooda, Vinita
DNA repair is more important than catalase for Salmonella virulence in mice.
Pathogenic microorganisms possess antioxidant defense mechanisms for protection from reactive oxygen metabolites such as hydrogen peroxide (H2O2), which are generated during the respiratory burst of phagocytic cells. These defense mechanisms include enzymes such as catalase, which detoxify reactive oxygen species, and DNA repair systems which repair damage resulting from oxidative stress. To determine the relative importance of these two potentially protective defense mechanisms against oxidative stress encountered by Salmonella during infection of the host, a Salmonella typhimurium double mutant unable to produce either the HPI or HPII catalase was constructed, and compared with an isogenic recA mutant deficient in DNA repair. The recA mutant was hypersusceptible to H2O2 at low cell densities in vitro, while the catalase mutant was more susceptible to high H2O2 concentrations at high cell densities. The catalase mutant was found to be resistant to macrophages and retained full murine virulence, in contrast to the recA mutant which previously was shown to be macrophage-sensitive and attenuated in mice. These observations suggest that Salmonella is subjected to low concentrations of H2O2 while at relatively low cell density during infection, conditions requiring an intact DNA repair system but not functional catalase activity. Images
Buchmeier, N A; Libby, S J; Xu, Y; Loewen, P C; Switala, J; Guiney, D G; Fang, F C
Relationship between uptake of mercury vapor by mushrooms and its catalase activity
The uptake of mercury vapor by mushrooms (Shiitake) artifically grown on an oak tree and the uptake in vitro by catalase extracts prepared from mushroom Hay Bacillus and spinach are reported. Mushrooms were exposed to 1.4 mg/Hg/cu m for 11 days. Measurement of total mercury was as previously described (Ogata et al. 1978, 1979). Levels in mushrooms ranged from 0.4 +/- 0.1 ..mu..g/g at 0.5 days to 4.6 +/- 0.2 ..mu..g/g at 10.5 days and steady-state thereafter. In in vitro studies Hy uptake by mushroom catalase extract was estimated by the perborate method. Uptake was found to parallel catalase activity and was inhibited by potassium cyanide, sodium azide, and 3-amino-1,2,4-triazole. Similar results were obtained with Hay Bacillus and spinach catalase extracts. Results suggest that the level of mercury in the mushroom can be used as an indicator of mercury pollution in the environment. It is also suggested that catalase has an important role in uptake of mercury vapor in the plant. 2 tables (JMT)
Ogata, M.; Kenmotsu, K.; Hirota, N.; Naito, M.
Development of a new catalase activity assay for biological samples using optical CUPRAC sensor.
A novel catalase activity assay was developed for biological samples (liver and kidney tissue homogenates) using a rapid and low-cost optical sensor-based 'cupric reducing antioxidant capacity' (CUPRAC) method. The reagent, copper(II)-neocuproine (Cu(II)-Nc) complex, was immobilized onto a cation-exchanger film of Nafion, and the absorbance changes associated with the formation of the highly-colored Cu(I)-Nc chelate as a result of reaction with hydrogen peroxide (H2O2) was measured at 450nm. When catalase was absent, H2O2 produced the CUPRAC chromophore, whereas catalase, being an effective H2O2 scavenger, completely annihilated the CUPRAC signal due to H2O2. Thus, the CUPRAC absorbance due to H2O2 oxidation concomitant with Cu(I)-Nc formation decreased proportionally with catalase. The developed sensor gave a linear response over a wide concentration range of H2O2 (0.68-78.6?M). This optical sensor-based method applicable to tissue homogenates proved to be efficient for low hydrogen peroxide concentrations (physiological and nontoxic levels) to which the widely used UV method is not accurately responsive. Thus, conventional problems of the UV method arising from relatively low sensitivity and selectivity, and absorbance disturbance due to gaseous oxygen evolution were overcome. The catalase findings of the proposed method for tissue homogenates were statistically alike with those of HPLC. PMID:24887508
Bekde?er, Burcu; Ozy�rek, Mustafa; G��l�, Kubilay; Alkan, Fulya �st�n; Apak, Re?at
Production of catalases by Aspergillus niger isolates as a response to pollutant stress by heavy metals
Isolates of Aspergillus niger, selected from the coal dust of a mine containing arsenic (As; 400 mg/kg) and from the river sediment of mine surroundings (As, 1651 mg/kg, Sb, 362 mg/kg), growing in minimal nitrate medium in the phase of hyphal development and spore formation, exhibited much higher levels of total catalase activity than the same species from the culture collection or a culture adapted to soil contaminated with As (5 mg/L). Electrophoretic resolution of catalases in cell-free extracts revealed three isozymes of catalases and production of individual isozymes was not significantly affected by stress environments. Exogenously added stressors (As{sup 5+}, Cd{sup 2+}, Cu{sup 2+}) at final concentrations of 25 and 50 mg/L and H{sub 2}O{sub 2} (20 or 40 m(M)) mostly stimulated production of catalases only in isolates from mines surroundings, and H{sub 2}O{sub 2} and Hg{sup 2+} caused the disappearance of the smallest catalase I. Isolates exhibited a higher tolerance of the toxic effects of heavy metals and H{sub 2}O{sub 2}, as monitored by growth, than did the strain from the culture collection.
Buckova, M.; Godocikova, J.; Simonovicova, A.; Polek, B. [Slovakian Academy of Science, Bratislava (Slovakia)
Production of catalases by Aspergillus niger isolates as a response to pollutant stress by heavy metals.
Isolates of Aspergillus niger, selected from the coal dust of a mine containing arsenic (As; 400 mg/kg) and from the river sediment of mine surroundings (As, 1651 mg/kg, Sb, 362 mg/kg), growing in minimal nitrate medium in the phase of hyphal development and spore formation, exhibited much higher levels of total catalase activity than the same species from the culture collection or a culture adapted to soil contaminated with As (5 mg/L). Electrophoretic resolution of catalases in cell-free extracts revealed three isozymes of catalases and production of individual isozymes was not significantly affected by stress environments. Exogenously added stressors (As(5+), Cd(2+), Cu(2+)) at final concentrations of 25 and 50 mg/L and H(2)O(2) (20 or 40 mM) mostly stimulated production of catalases only in isolates from mines surroundings, and H(2)O(2) and Hg(2+) caused the disappearance of the smallest catalase I. Isolates exhibited a higher tolerance of the toxic effects of heavy metals and H(2)O(2), as monitored by growth, than did the strain from the culture collection. PMID:15902463
Buckov�, Maria; Godoc�kov�, Jana; Simonovicov�, Alexandra; Polek, Bystr�k
Characterization of a Facultatively Psychrophilic Bacterium, Vibrio rumoiensis sp. nov., That Exhibits High Catalase Activity
A novel facultatively psychrophilic bacterium, strain S-1, which exhibits extraordinarily high catalase activity was isolated from the drain pool of a fish product processing plant that uses H2O2 as a bleaching and microbicidal agent. The catalase activity of the isolate was 1 or 2 orders of magnitude higher than those of Corynebacterium glutamicum, Staphylococcus aureus, Pseudomonas fluorescens, and five other species tested in this study. The strain seemed to possess only one kind of catalase, according to the results of polyacrylamide gel electrophoresis of the cell extract. The optimum temperature for catalase activity was about 30�C, which was about 20�C lower than that for bovine catalase activity. Electron microscopic observation revealed that the surface of the microorganism was covered by blebs. Although the isolate was nonflagellated, its taxonomic position on the basis of physiological and biochemical characteristics and analysis of 16S rRNA sequence and DNA-DNA relatedness data indicated that strain S-1 is a new species belonging to the genus Vibrio. Accordingly, we propose the name Vibrio rumoiensis. The type strain is S-1 (FERM P-14531).
Yumoto, Isao; Iwata, Hideaki; Sawabe, Tomoo; Ueno, Keisuke; Ichise, Nobutoshi; Matsuyama, Hidetoshi; Okuyama, Hidetoshi; Kawasaki, Kosei
Differential expression of three catalase genes in the small radish (Rhaphanus sativus L. var. sativus).
Three catalase cDNA clones were isolated from the small radish (Raphanus sativus L.). Their nucleotide and deduced amino acid sequences showed the greatest homology to those of Arabidopsis. Genomic Southern blot analysis, using RsCat1 cDNA as a probe, showed that catalases are encoded by small multigene family in the small radish. Nondenaturing polyacrylamide gels revealed the presence of several catalase isozymes, the levels of which varied among the organs examined. The isozyme activities were assigned the individual catalase genes by Northern analysis using total RNA from different organs. The three catalase genes were differentially expressed in response to treatments such as white light, xenobiotics, osmoticum, and UV. Their expression in seedlings was controlled by the circadian clock under a light/dark cycle and/or in constant light. Interestingly, RsCat1 transcripts peaked in the morning, while those of RsCat2 and RsCat3 peaked in the early evening. Our results suggest that the RsCat enzymes are involved in defense against the oxidative stress induced by environmental changes. PMID:17846497
Kwon, Soon Il; Lee, Hyoungseok; An, Chung Sun
Antifungal Effect of Hydrogen Peroxide on Catalase-Producing Strains of Candida spp.
Objective: Clinical isolates of Candida were tested for the presence of catalase and susceptibility to hydrogen peroxide. Methods: MIC was tested by broth dilution technique and catalase was determined by a spectrophotometric procedure. Results: All 38 strains tested were inhibited by hydrogen peroxide in concentrations ranging from 4.4 to 88 mM/l, with non-albicans isolates generally requiring higher concentrations of hydrogen peroxide for inhibition. Growth media consisting of glucose and protein diminished the antifungal effectiveness of hydrogen peroxide, as did the presence of hemoglobin, in incubation mixtures. However, hydrogen peroxide exerted greater inhibition at pH 4 than at pH 7. Although all Candida isolates tested possessed catalase, there was no apparent correlation between the catalase activity of individual isolates and the minimal antifungal concentration of hydrogen peroxide. Conclusions: This study suggested that, despite the production of catalase by vaginal microorganisms, hydrogen peroxide may exert a regulating influence which may be further modified by the proteins found in the vaginal milieu.
White, Sandra
Silencing an Anopheles gambiae catalase and sulfhydryl oxidase increases mosquito mortality after a blood meal.
Catalase is a potent antioxidant, likely involved in post-blood meal homeostasis in mosquitoes. This enzyme breaks down H2O2, preventing the formation of the hydroxyl radical (HO*). Quiescins are newly classified sulfhydryl oxidases that bear a thioredoxin motif at the N-terminal and an ERV1-like portion at the C-terminal. These proteins have a major role in generating disulfides in intra- or extracellular environments, and thus participate in redox reactions. In the search for molecules to serve as targets for novel anti-mosquito strategies, we have silenced a catalase and a putative quiescin/sulfhydryl oxidase (QSOX), from the African malaria vector Anopheles gambiae, through RNA interference (RNAi) experiments. We observed that the survival of catalase- and QSOX-silenced insects was reduced over controls following blood digestion, most likely due to the compromised ability of mosquitoes to scavenge and/or prevent damage caused by blood meal-derived oxidative stress. The higher mortality effect was more accentuated in catalase-silenced mosquitoes, where catalase activity was reduced to low levels. Lipid peroxidation was higher in QSOX-silenced mosquitoes suggesting the involvement of this protein in redox homeostasis following a blood meal. This study points to the potential of molecules involved in antioxidant response and redox metabolism to serve as targets of novel anti-mosquito strategies and offers a screening methodology for finding targetable mosquito molecules. PMID:18454489
Magalhaes, T; Brackney, D E; Beier, J C; Foy, B D
Silencing an Anopheles gambiae Catalase and Sulfhydryl Oxidase Increases Mosquito Mortality After a Blood Meal
Catalase is a potent antioxidant, likely involved in post-blood meal homeostasis in mosquitoes. This enzyme breaks down H2O2, preventing the formation of the hydroxyl radical (HO�). Quiescins are newly classified sulfhydryl oxidases that bear a thioredoxin motif at the N-terminal and an ERV1-like portion at the C-terminal. These proteins have a major role in generating disulfides in intra- or extracellular environments, and thus participate in redox reactions. In the search for molecules to serve as targets for novel anti-mosquito strategies, we have silenced a catalase and a putative quiescin/sulfhydryl oxidase (QSOX), from the African malaria vector Anopheles gambiae, through RNA interference (RNAi) experiments. We observed that the survival of catalase- and QSOX-silenced insects was reduced over controls following blood digestion, most likely due to the compromised ability of mosquitoes to scavenge and/or prevent damage caused by blood meal-derived oxidative stress. The higher mortality effect was more accentuated in catalase-silenced mosquitoes, where catalase activity was reduced to low levels. Lipid peroxidation was higher in QSOX-silenced mosquitoes suggesting the involvement of this protein in redox homeostasis following a blood meal. This study points to the potential of molecules involved in antioxidant response and redox metabolism to serve as targets of novel anti-mosquito strategies and offers a screening methodology for finding targetable mosquito molecules.
Magalhaes, T.; Brackney, D.E.; Beier, J.C.; Foy, B.D.
Catalase is a sink for H2O2 and is indispensable for stress defence in C3 plants
Hydrogen peroxide (H2O2) has been implicated in many stress conditions. Control of H2O2 levels is complex and dissection of mechanisms generating and relieving H2O2 stress is difficult, particularly in intact plants. We have used transgenic tobacco with ?10% wild-type catalase activity to study the role of catalase and effects of H2O2 stress in plants. Catalase-deficient plants showed no visible disorders
Hilde Willekens; Sangpen Chamnongpol; Mark Davey; Martina Schraudner; Christian Langebartels; Dirk Inz�; Wim Van Camp; Marc Van Montagu
[Study of catalase and proteolytic activities of different variants of Bacillus mesentericus].
Catalase and proteolytic activity of the culures and morphological variants of Bacillus mesentericus fuscus, Bac. mesentericus vulgatus were studied. The variants were obtained as a result of prolonged cultivation of the stock strains in the potato mash under the layer of vaseline oil. The level of catalase activity varies in different morphological variants of the same culture, changes with age and depends on the storage conditions. The catalase activity in the rough, smooth and papillar variants that were freshly isolated from the potato mash was 1.5=2.5 times lower than that in the variants long kept on the agar medium. The quantitative indexes of the proteolytic activity of different variants also varied. PMID:1208411
Vasilevskaya, I A; Kolchinskaya, I D; Sergeichuk, M G; Roy, A A
Influence of Catalase and Superoxide Dismutase on Ozone Inactivation of Listeria monocytogenes
The effects of ozone at 0.25, 0.40, and 1.00 ppm on Listeria monocytogenes were evaluated in distilled water and phosphate-buffered saline. Differences in sensitivity to ozone were found to exist among the six strains examined. Greater cell death was found following exposure at lower temperatures. Early stationary-phase cells were less sensitive to ozone than mid-exponential- and late stationary-phase cells. Ozonation at 1.00 ppm of cabbage inoculated with L. monocytogenes effectively inactivated all cells after 5 min. The abilities of in vivo catalase and superoxide dismutase to protect the cells from ozone were also examined. Three listerial test strains were inactivated rapidly upon exposure to ozone. Both catalase and superoxide dismutase were found to protect listerial cells from ozone attack, with superoxide dismutase being more important than catalase in this protection.
Fisher, Christopher W.; Lee, Dongha; Dodge, Beth-Anne; Hamman, Kristen M.; Robbins, Justin B.; Martin, Scott E.
The catalase-hydrogen peroxide system. Kinetics of catalatic action at high substrate concentrations
1. A re-examination of the catalase�hydrogen peroxide reaction at high substrate concentrations, by using the quenched-flow technique, reveals a more complex kinetic behaviour than that previously reported. At constant reaction time the catalatic process obeys Michaelis�Menten kinetics, but the apparent Michaelis constant is markedly time-dependent, whereas the conventional catalase activity is independent of time. 2. The kinetics of the `time effect' were analysed and it is suggested that the effect derives from the formation of an inactive species (thought to be catalase Compound II). The process shows Michaelis�Menten kinetics, with a Michaelis constant equal to that for the catalatic reaction in the limit of zero reaction time. 3. It has been confirmed that certain buffer components have marked inhibitory effects on the catalatic reaction and that, in unbuffered systems, catalatic activity is substantially independent of pH in the range 4�7�10�5.
Jones, Peter; Suggett, A.
The functionalization of saturated hydrocarbons. Part 35. On the intermediates in an Fe III catalase model in pyridine. Relevance to the catalase enzyme
Ferric chloride in pyridine behaves as an efficient model for the catalase enzyme. It converts H2O2 nearly quantitatively into water and oxygen (2 H2O2 ? 2 H2O + O2). The addition of Ph2S to the model system affords Ph2SO, the amount of which increases with the Ph2S added. The inverse relationship between oxygen and Ph2SO formation proves that there is
Derek H. R. Barton; Bin Hu
Vascular endothelium-specific overexpression of human catalase in cloned pigs
The objective of this study was to develop transgenic Yucatan minipigs that overexpress human catalase (hCat) in an endothelial-specific manner. Catalase metabolizes hydrogen peroxide (H2O2), an important regulator of vascular tone that contributes to diseases such as atherosclerosis and preeclampsia. A large animal model to study reduced endothelium-derived H2O2 would therefore generate valuable translational data on vascular regulation in health and disease. Yucatan minipig fetal fibroblasts stably co-transfected with human catalase (Tie2-hCat) and eGFP expression constructs were isolated into single-cell populations. The presence of the Tie2-hCat transgene in individual colonies of fibroblasts was determined by PCR. Transgenic fibroblasts were used for nuclear transfer into enucleated oocytes by electrofusion. A minimum of 140 cloned embryos were transferred per surrogate sow (n = 4). All four surrogates maintained pregnancies and piglets were delivered by cesarean section. Nine male piglets from three of the four litters carried the Tie2-hCat transgene. Expression of human catalase mRNA and overall elevated catalase protein in isolated umbilical endothelial cells from transgenic piglets were verified by RT�PCR and western blot, respectively, and endothelial localization was confirmed by immunohistochemistry. Increased enzymatic activity of catalase in transgenic versus wild-type endothelial cells was inferred based on significantly reduced levels of H2O2 in culture. The similarities in swine and human cardiovascular anatomy and physiology will make this pig model a valuable source of information on the putative role of endothelium-derived H2O2 in vasodilation and in the mechanisms underlying vascular health and disease.
Samuel, M.; Mahan, E.; Padilla, J.; Simmons, G. H.; Arce-Esquivel, A. A.; Bender, S. B.; Whitworth, K. M.; Hao, Y. H.; Murphy, C. N.; Walters, E. M.; Prather, R. S.; Laughlin, M. H.
Coordination modes of bridge carboxylates in dinuclear manganese compounds determine their catalase-like activities.
To explore the role of bridge carboxylate coordination modes on the catalase-like activities of dinuclear manganese compounds, [Mn(II)2(bpmapa)2(H2O)2](ClO4)2 (1), [Mn(II)2(pbpmapa)2(H2O)2](ClO4)2 (2), and [Mn(II)2(bpmaa)2(H2O)3](ClO4)2 (3) (bpmapa = [bis(2-pyridylmethyl)amino]propionic acid, pbpmapa = alpha-phenyl-beta-[bis(2-pyridylmethyl)amino]propionic acid, and bpmaa = [bis(2-pyridylmethyl)amino]acetic acid), in which Mn(II)-Mn(II) centers have a similar coordination sphere but different carboxylate-Mn bridging modes have been synthesized and structurally characterized by single X-ray diffraction, UV-visible, IR, and EPR spectroscopies, and their catalase-like activities were investigated. Studies of their catalytic activities and the influence of the nitrogenous bases on their catalytic activities indicated that the carboxylate-Mn coordination mode was crucial in H2O2 deprotonation, and eventually in H2O2 disproportionation. Compound 1 with a bidentate carboxylate bridge showed higher catalase-like activity than 2 and 3, in which the carboxylate groups have a monodentate bridging mode. The deprotonation ability of the carboxylate anion was determined by the O-C-O angle and the distance between the weakly bound oxygen of the bridging carboxylate to the manganese ion. The smaller the angle, and the shorter the distance, the stronger the basicity that the carboxylate anion exhibits. The bidentate mu-1,1 bridging coordination mode functionally mimicked the glutamate residues at the manganese catalase active site. Our results suggested that increasing the basicity of the bridging carboxylate ligand of the catalase model compounds will increase their deprotonation ability and lead to more active catalase mimics. PMID:19809747
Jiang, Xiaojun; Liu, Hui; Zheng, Bing; Zhang, Jingyan
Catalase-like catalytic reaction of the dinuclear manganese�salen complex
Catalase-like activity of a dinuclear manganese-salen (Mn�salen) complex, [Mn(salen)(H2O)]2(ClO4)2 (salen = N,N ?-bis(salicylidene)-1,2-diaminoethane), was investigated. The dinuclear Mn�salen complex exhibits higher catalase-like activity than that of the mononuclear Mn�salen compound, and its activity can be enhanced by an external base. Different reaction intermediates in the presence and absence of an external base were observed, and the catalytically active species was
Rong Li; Fuping Huang; Xiaojun Jiang; Mingyuan Liu; Yanying Song; Hui Liu; Jingyan Zhang
Polyaniline based catalase biosensor for the detection of hydrogen peroxide and azide
The CAT\\/PANi\\/ITO bioelectrode has been prepared as a catalase biosensor and shows response for monitoring not only of H2O2 but also azide. The sensor exhibited an excellent response to the H2O2 and azide. The linear range of H2O2 was 0.064?1 mM and for azide 0.125?4 mM, respectively. Catalase biosensor was based on the principle of the measurements\\u000a as the decrease
Ravindra P. Singh; Da-Yeon Kang; Byung-Keun Oh; Jeong-Woo Choi
[Activity of NAD- and NADP-containing enzymes and catalase in human erythrocytes after sucrose loading].
The dynamics of glucose content and activity of GL-6-FDG, MDG, ICDG and of catalase in the erythrocytes of healthy people under glucose load was investigated. It has been established that maximal increase of the glucose content in blood under glucose load occurs 60 min later and the peak of activity of all the studied enzymes--90 min later. A degree of the activity increase in certain enzymes is not the same. It enhances considerably in GL-6-FDG and catalase and is hardly tracable in MDG and ICDG. A conclusion is made that glucose metabolism in erythrocytes is accompanied by the intensification of synthesis and hydrogen peroxide decomposition processes. PMID:3810896
Storozhuk, P G; Skliar, V A; Korochanskaia, S P; Bykov, I M
The effect of intracellular antioxidant delivery (catalase) on hydrogen peroxide and proinflammatory cytokine synthesis: a new therapeutic horizon.
Reactive oxygen species synthesized by endothelial cells may be responsible for cell damage and altered physiologic function. After endotoxin stimulation, free radicals including H(2)O(2) are produced. We have developed a method of intracellular drug delivery using albumin microcapsules. Catalase would be an excellent compound to alter H(2)O(2) production. However, the large molecular size of catalase limits cellular penetration. Endothelial cells have been previously shown to readily phagocytoze albumin microcapsules. Methods: Catalase was added to an albumin solution to form a 10% solution of catalase. Microspheres from 2 to 7 microm in size were formed using a Bucchi spray dryer. Human endothelial cells were incubated with varying concentrations of microencapsulated catalase. The cells were then exposed to Escherichia coli endotoxin to determine if increased intracellular penetration of catalase would inhibit H(2)O(2), nitrate, and cytokine synthesis. Results: There was a 7.2-fold increase in endothelial intracellular catalase after 48 h incubation. H(2)O(2) was inhibited by 72%, nitrate 96%, TNF 90%, IL1 21%, IL6 42%. Conclusions: These results demonstrate that inhibition of H(2)O(2) as a result of increased intracellular delivery of catalase inhibits proinflammatory cytokine synthesis after endotoxin exposure. PMID:19845487
Siwale, Rodney C; Yeboah, George K; Addo, Richard; Oettinger, Carl W; D'Souza, Martin J
Morphological transformation and catalase activity of Syrian hamster embryo cells treated with hepatic peroxisome proliferators, TPA and nickel sulphate.
The abilities of the hepatic peroxisome proliferators (HPPs) clofibrate, di(2-ethylhexyl)phthalate (DEHP), mono(2-ethylhexyl)-phthalate (MEHP), 2,4-dichlorophenoxy acetic acid (2,4-D), 2,4,5-trichlorophenoxy acetic acid (2,4,5-T) and tiadenol to induce morphological transformation and to increase the catalase activity of Syrian hamster embryo (SHE) cells were studied. DEHP, MEHP, clofibrate and tiadenol induced morphological transformation of SHE cells and increased the catalase activity. DEHP was more potent than clofibrate and tiadenol in both inducing catalase and morphological transformation, while MEHP seemed more potent than DEHP in inducing catalase, but not morphological transformation, 2,4,5-T and 2,4-D did not induce morphological transformation, but 2,4,5-T was more potent than clofibrate in increasing the catalase activity. These results show that several HPPs induce morphological transformation of SHE cells and an increase in the catalase activity. There is, however, no direct connection between these two parameters, as seen from the results of 2,4,5-T. The tumor promoter TPA, and the metal salt nickel sulphate, induced morphological transformation of SHE cells without any appreciable increase in the catalase activity. These results further corroborate the dissociation between induction of morphological transformation and the increase in catalase activity. PMID:2334865
Mikalsen, S O; Holen, I; Sanner, T
Cytochemical localization of catalase and several hydrogen peroxide-producing oxidases in the nucleoids and matrix of rat liver peroxisomes
Synopsis��The distribution of catalase, amino acid oxidase, -hydroxy acid oxidase, urate oxidase and alcohol oxidase was studied cytochemically in rat hepatocytes. The presence of catalase was demonstrated with the conventional diaminobenzidine technique. Oxidase activities were visualized with methods based on the enzymatic or chemical trapping of the hydrogen peroxide produced by these enzymes during aerobic incubations.All enzymes investigated were found
M. Veenhuis; S. E. Wendelaar Bonga
The importance of carboxylate ligands in the differentiation of catalase reactivity from Gif ketonization systems
The presence or absence of certain chelating carboxylic acids such as picolinic acid permits the distinction between ketonization (Gif Chemistry) and oxygen formation (catalase reaction). In the presence of such an acid, evidence is provided for the possible involvement of a IIIFe?O?O?FeIII species as a key intermediate in this hydrocarbon activation chemistry.
Derek H. R Barton; Bin Hu; Dennis K Taylor; Roy U Rojas Wahl
Catalase-like activity of a water-soluble complex of Ru(II)
A Ru(II) complex of an unsymmetrical macrocycle (containing two tertiary amine nitrogen atoms acting as ligands) exhibits a marked catalytic activity for disproportionation of hydrogen peroxide, with both high rate and turnover number. Although the stoichiometry is the same as for catalase, the reaction does not obey the simple kinetic law reported for this enzyme. The proposed reaction scheme includes
S. Choua; P. Pacheco; C. Coquelet; E. Bienven�e
Catalase-like oxygen production by horseradish peroxidase must predominantly be an enzyme-catalyzed reaction.
When hydrogen peroxide (H2O2) was provided as the only substrate for horseradish peroxidase C (HRP-C) the catalase-like emission of oxygen gas was observed. The reaction was favored at neutral compared to acidic pH. Addition of the superoxide radical scavengers tetranitromethane (TNM) or superoxide dismutase (SOD) increased activity. TNM's effect was concentration dependent but SOD's was not, indicating that only some of the superoxide generated was released into solution. Manganous ions (Mn2+) react with superoxide radicals to regenerate H2O2 but not oxygen; when added to the reaction medium oxygen production was reduced but not abolished. The effect was essentially concentration independent, suggesting that most oxygen was produced enzymatically and not by chemical disproportionation of superoxide. The catalase-like activities of some site-directed mutants of HRP-C suggest that active site residues histidine 42 and arginine 38 are influential in determining this activity. A clear correlation also existed between catalase activity and the enzymes' resistance to inactivation by H2O2. Computer simulation of a reaction scheme that included catalase-like activity agreed well with experimental data. PMID:11488605
Hiner, A N; Hern�ndez-Ruiz, J; Williams, G A; Arnao, M B; Garc�a-C�novas, F; Acosta, M
Impedance spectroscopy and conductometric biosensing for probing catalase reaction with cyanide as ligand and inhibitor.
In this work, a new biosensor was prepared through immobilization of bovine liver catalase in a photoreticulated poly (vinyl alcohol) membrane at the surface of a conductometric transducer. This biosensor was used to study the kinetics of catalase-H(2)0(2) reaction and its inhibition by cyanide. Immobilized catalase exhibited a Michaelis-Menten behaviour at low H(2)0(2) concentrations (<100mM) with apparent constant K(M)(app)=84�3mM and maximal initial velocity V(M)(app)=13.4?S min(-1). Inhibition by cyanide was found to be non-competitive and inhibition binding constant K(i) was 13.9�0.3?M. The decrease of the biosensor response by increasing cyanide concentration was linear up to 50?M, with a cyanide detection limit of 6?M. In parallel, electrochemical characteristics of the catalase/PVA biomembrane and its interaction with cyanide were studied by cyclic voltammetry and impedance spectroscopy. Addition of the biomembrane onto the gold electrodes induced a significant increase of the interfacial polarization resistance R(P). On the contrary, cyanide binding resulted in a decrease of Rp proportional to KCN concentration in the 4 to 50?M range. Inhibition coefficient I(50) calculated by this powerful label-free and substrate-free technique (24.3?M) was in good agreement with that determined from the substrate-dependent conductometric biosensor (24.9?M). PMID:20813591
Bouyahia, Naima; Hamlaoui, Mohamed Larbi; Hnaien, Mouna; Lagarde, Florence; Jaffrezic-Renault, Nicole
Effect of kinetin on growth, auxin catabolism, peroxidase and catalase activities
Kinetin causes an inhibition of growth of root sections. The IAA-oxidizing system from treated roots is activated. Kinetin parallely produces an increase in peroxidase activity but does not affect catalase. The compound has no effect in vitro on the enzymes considered.
Thomas Gaspar; Anne Xhaufflaire
Antioxidant Effect of Licorice Root on Blood Catalase Activity in Vibration Stress
Rabbits were treated (orally) with a preparation of Glycyrrhiza glabra L. for 30 days and in parallel were exposed to vibration stress (30 days). The licorice preparation reduced catalase activity in the peripheral blood and increased animal resistance to vibration stress.
K. R. Oganesyan
Enhancement of the nitrogen fixation efficiency of genetically-engineered Rhizobium with high catalase activity.
The vktA catalase gene, which had been cloned from Vibrio rumoiensis S-1T having extraordinarily high catalase activity, was introduced into the root nodule bacterium, Rhizobium leguminosarum bv. phaseoli USDA 2676. The catalase activity of the vktA-transformed R. leguminosarum cells (free-living) was three orders in magnitude higher than that of the parent cells and this transformant could grow in a higher concentration of exogenous hydrogen peroxide (H2O2). The vktA-transformant was inoculated to the host plant (Phaseolus vulgaris L.) and the nodulation efficiency was evaluated. The results showed that the nitrogen-fixing activity of nodules was increased 1.7 to 2.3 times as compared to the parent. The levels of H2O2 in nodules formed by the vktA-transformant were decreased by around 73%, while those of leghemoglobins (Lba and Lbb) were increased by 1.2 (Lba) and 2.1 (Lbb) times compared with the parent. These results indicated that the increase of catalase activity in rhizobia could be useful to improve the nitrogen-fixing efficiency of nodules by the reduction of H2O2 content concomitantly with the enhancement of leghemoglobins contents. PMID:20547375
Orikasa, Yoshitake; Nodasaka, Yoshinobu; Ohyama, Takuji; Okuyama, Hidetoshi; Ichise, Nobutoshi; Yumoto, Isao; Morita, Naoki; Wei, Min; Ohwada, Takuji
Catalase and superoxide dismutase activities in a Stenotrophomonas maltophilia WZ2 resistant to herbicide pollution.
Quinclorac bensulfuron-methyl is a mixed herbicide widely used on paddy rice field to effectively control barnyard grass and most broad-leaved grasses and sedges. We analyzed superoxide dismutase (SOD) and catalase activities in the quinclorac-highly degrading strain Stenotrophomonas maltophilia WZ2 and Gram-negative standard strain Escherichia coli K12 in an attempt to understand antioxidant enzymes in bacteria are produced in response to quinclorac or bensulfuron-methyl, which increases the virulence of the bacteria. MnSOD and two additional catalase isozymes were induced by quinclorac or bensulfuron-methyl in S. maltophilia WZ2, but not in E. coli K12. Quinclorac turned out to be a more sensitive inducer of SOD, whereas bensulfuron-methyl is a more sensitive one of catalase. A mixture of both has effects similar to quinclorac. Results indicate that catalase has a much weakly role in the defense against quinclorac or bensulfuron-methyl induced oxidative stress, whereas SOD could be critical. PMID:18304632
L�, Zhenmei; Sang, Liya; Li, Zimu; Min, Hang
Antioxidant enzymes in cabbage: Variability and inheritance of superoxide dismutase, peroxidase and catalase
Cruciferous vegetables are important source of dietary nutrients and antioxidants. Antioxidants have been touted as beneficial for enhancing plant stand and mitigating the effects of biotic and abiotic stresses. The objective of the present study was to determine variability for superoxide dismutase, peroxidase and catalase activity; its transmissibility; and correlation. Samples were harvested at fresh market stage, frozen immediately in
B. K. Singh; S. R. Sharma
Cloning and expression of the catalase gene from the anaerobic bacterium Desulfovibrio vulgaris (Miyazaki F).
We identified a gene encoding a catalase from the anaerobic bacteria Desulfovibrio vulgaris (Miyazaki F), and the expression of its gene in Escherichia coli. The 3.3-kbp DNA fragment isolated from D. vulgaris (Miyazaki F) by double digestion with EcoRI and SalI was found to produce a protein that binds protoheme IX as a prosthetic group in E. coli. This DNA fragment contained a putative open reading frame (Kat) and one part of another open reading frame (ORF-1). The amino acid sequence of the amino terminus of the protein purified from the transformed cells was consistent with that deduced from the nucleotide sequence of Kat in the cloned fragment of D. vulgaris (Miyazaki F) DNA, which may include promoter and regulatory sequences. The nucleotide sequence of Kat indicates that the protein is composed of 479 amino acids per monomer. The recombinant catalase was found to be active in the decomposition of hydrogen peroxide, as are other catalases from aerobic organisms, but its K(m) value was much greater. The hydrogen peroxide stress against D. vulgaris (Miyazaki F) induced the activity for the decomposition of hydrogen peroxide somewhat, so the catalase gene may not work effectively in vivo. PMID:11226874
Kitamura, M; Nakanishi, T; Kojima, S; Kumagai, I; Inoue, H
Sunflower cotyledons cope with copper stress by inducing catalase subunits less sensitive to oxidation
Copper is an essential trace element for living organisms, in excess, can be toxic to the cell because of its capacity to generate reactive oxygen species (ROS). Catalase (CAT) catalyzes the dismutation of hydrogen peroxide into water and dioxygen and in plants it is located in peroxisomes and glyoxysomes. Different metals can induce changes in CAT activity, but the mechanism
Liliana B. Pena; Claudia E. Azpilicueta; Susana M. Gallego
Do Superoxide Dismutase (SOD) and Catalase (CAT) protect Cells from DNA Damage Induced by Active Arsenicals?
Superoxide dismutase (SOD) catalyzes the conversion of superoxide to hydrogen peroxide, which can be converted to water and oxygen through the action of catalase. Heterozygous mice of strain B6: 129S7-SodltmlLeb/J were obtained from Jackson Laboratories and bred to produce offspr...
Assembly of catalase-based bioconjugates for enhanced anticancer efficiency of photodynamic therapy in vitro.
An oxygen generation core-shell structure uploading rose bengal has been fabricated by covalent assembly of catalase and alginate dialdehyde via Schiff's base. The composite can catalyze the decomposition of intracellular H2O2 to increase the concentration of O2, which effectively enhances the anticancer efficiency of photodynamic therapy in vitro. PMID:24104860
Zhao, Jie; Fei, Jinbo; Du, Cuiling; Cui, Wei; Ma, Hongchao; Li, Junbai
In a previous report ( 1930) upon the respiratory exchange and cata lase in tissues of lamellibranchs, the author pointed out a relationship between the habitat distribution of several species and the relative abun dance of their muscle catalase. To these have been added new data for the same '�? and for other species on our southern Atlantic coast, making
Catalase activity and expression in developing sunflower seeds as related to drying
Changes in catalase (CAT) activity and in CAT iso- form pattern and expression were investigated in developing sunflower (Helianthus annuus L.) seeds during desiccation on the mother plant and after arti- ficial drying on the flowerheads. Seeds regularly des- iccated during their development on the mother plant and reached mass maturity at c. 42 d after flowering (DAF). Freshly harvested
Christophe Bailly; Juliette Leymarie; Arnaud Lehner; Sandra Rousseau; Francoise Corbineau
Catalase and superoxide dismutase in alfalfa root nodules. [Medicago sativa L
Catalase and superoxide dismutase (SOD), in scavenging H/sub 2/ O/sub 2/ and O/sub 2/, respectively, have been recently proposed to play a role in leghemoglobin protection. The occurrence of catalase and SOD activities in alfalfa (Medicago sativa L.) nodule cytosol is reported here. Enzymes were extracted at 0-4/sup 0/C from 0.5 g fresh nodules with 12 ml of a medium containing K-phosphate buffer 50 mM, pH 7.8 and Na/sub 2/EDTA 0.1 mM. The homogenate was filtered and centrifuged at 18,000 xg for 10 min, and the resulting supernatant was used for catalase assay. A further precipitation of leghemoglobin was required to avoid interferences with SOD determination. Catalase was determined by back-titration with KMnO/sub 4/. SOD was assayed by measuring the inhibition of nitro blue tetrazolium reduction. The sensitivity of SOD activity to CN/sup -/ was tested by including 1 mM KCN in the reaction mixture. Catalase activity of alfalfa nodule cytosol was 237 +/- 1 units/mg protein, decreasing very significantly (P < 0.01, Duncan's multiple range test) at 20 mM NO/sub 3//sup -/. Typical specific SOD activities were 94 +/- 5 and 65 +/- 4 units/mg protein, without CN/sup -/ and with CN/sup -/, respectively. Both activities increased very significantly at 20 mM NO/sub 3//sup -/. SOD activities with CN/sup -/ were 70-80% those without CN/sup -/ within the range of NO/sub 3//sup -/ investigated (0-20 mM).
Becana, M.; Aparicio-Tejo, P.M.; Sanchez-Diaz, M.
Catalase Overexpression Fails to Attenuate Allergic Airways Disease in the Mouse1
Oxidative stress is a hallmark of asthma, and increased levels of oxidants are considered markers of the inflammatory process. Most studies to date addressing the role of oxidants in the etiology of asthma were based on the therapeutic administration of low m.w. antioxidants or antioxidant mimetic compounds. To directly address the function of endogenous hydrogen peroxide in the pathophysiology of allergic airway disease, we comparatively evaluated mice systemically overexpressing catalase, a major antioxidant enzyme that detoxifies hydrogen peroxide, and C57BL/6 strain matched controls in the OVA model of allergic airways disease. Catalase transgenic mice had 8-fold increases in catalase activity in lung tissue, and had lowered DCF oxidation in tracheal epithelial cells, compared with C57BL/6 controls. Despite these differences, both strains showed similar increases in OVA-specific IgE, IgG1, and IgG2a levels, comparable airway and tissue inflammation, and identical increases in procollagen 1 mRNA expression, following sensitization and challenge with OVA. Unexpectedly, mRNA expression of MUC5AC and CLCA3 genes were enhanced in catalase transgenic mice, compared with C57BL/6 mice subjected to Ag. Furthermore, when compared with control mice, catalase overexpression increased airway hyperresponsiveness to methacholine both in naive mice as well as in response to Ag. In contrast to the prevailing notion that hydrogen peroxide is positively associated with the etiology of allergic airways disease, the current findings suggest that endogenous hydrogen peroxide serves a role in suppressing both mucus production and airway hyperresponsiveness.
Reynaert, Niki L.; Aesif, Scott W.; McGovern, Toby; Brown, Amy; Wouters, Emiel F. M.; Irvin, Charles G.; Janssen-Heininger, Yvonne M. W.
Molecular cloning and nucleotide sequence of full-length cDNA for sweet potato catalase mRNA.
A nearly full-length cDNA clone for catalase (pCAS01) was obtained through immunological screening of cDNA expression library constructed from size-fractionated poly(A)-rich RNA of wounded sweet potato tuberous roots by Escherichia coli expression vector-primed cDNA synthesis. Two additional catalase cDNA clones (pCAS10 and pCAS13), which contained cDNA inserts slightly longer than that of pCAS01 at their 5'-termini, were identified by colony hybridization of another cDNA library. Those three catalase cDNAs contained primary structures not identical, but closely related, to one another based on their restriction enzyme and RNase cleavage mapping analyses, suggesting that microheterogeneity exists in catalase mRNAs. The cDNA insert of pCAS13 carried the entire catalase coding capacity, since the RNA transcribed in vitro from the cDNA under the SP6 phage promoter directed the synthesis of a catalase polypeptide in the wheat germ in vitro translation assay. The nucleotide sequencing of these catalase cDNAs indicated that 1900-base catalase mRNA contained a coding region of 1476 bases. The amino acid sequence of sweet potato catalase deduced from the nucleotide sequence was 35 amino acids shorter than rat liver catalase [Furuta, S., Hayashi, H., Hijikata, M., Miyazawa, S., Osumi, T. & Hashimoto, T. (1986) Proc. Natl Acad. Sci. USA 83, 313-317]. Although these two sequences showed only 38% homology, the sequences around the amino acid residues implicated in catalytic function, heme ligand or heme contact had been well conserved during evolution. PMID:2885193
Sakajo, S; Nakamura, K; Asahi, T
Oral administration of a catalase-producing Lactococcus lactis can prevent a chemically induced colon cancer in mice.
Reactive oxygen species, such as hydrogen peroxide (H2O2), are involved in various aspects of tumour development. Decreasing their levels can therefore be a promising approach for colon cancer prevention. The objective of this study was to evaluate the effect of catalase-producing Lactococcus lactis on the prevention of an experimental murine 1,2-dimethylhydrazine (DMH)-induced colon cancer. DMH-treated BALB/c mice received either a catalase-producing L. lactis strain or the isogenic non-catalase-producing strain as a control, whereas other untreated mice did not receive bacterial supplementation. Catalase activity and H2O2 levels in intestinal fluids and blood samples were measured, and changes in the histology of the large intestines during tumour progression were evaluated. The catalase-producing L. lactis strain used in this study was able to slightly increase catalase activities in DMH-treated mice (1.19+/-0.08 U ml(-1)) and reduce H2O2 levels (3.4+/-1.1 microM) compared to (i) animals that received the non-catalase-producing strain (1.00+/-0.09 U ml(-1), 9.0+/-0.8 microM), and (ii) those that did not receive bacterial supplementation (1.06+/-0.07 U ml(-1), 10.0+/-1.1 microM). Using the histopathological grading scale of chemically induced colorectal cancer, animals that received the catalase-producing L. lactis had a significantly lesser extent of colonic damage and inflammation (2.0+/-0.4) compared to animals that received the non-catalase-producing L. lactis (4.0+/-0.3) or those that did not receive bacterial supplementation (4.7+/-0.5). The catalase-producing L. lactis strain used in this study was able to prevent tumour appearance in an experimental DMH-induced colon cancer model. PMID:18065674
de Moreno de LeBlanc, Alejandra; LeBlanc, Jean Guy; Perdig�n, Gabriela; Miyoshi, Anderson; Langella, Philippe; Azevedo, Vasco; Sesma, Fernando
Resonance Scattering Spectral Detection of Catalase Activity Using Au@Ag Nanoparticle as Probe and Coupling Catalase Catalytic Reaction with Fenton Reaction
The AucoreAgshell (Au@Ag) nanoparticles in size of 30�nm were prepared using 10�nm gold nanoparticles as seeds at 90�C, and were purified by\\u000a high-speed centrifugation to remove the excess trisodium citrate to obtain Au@Ag nanoprobe. In the medium of pH�4.0 acetate\\u000a buffer solution�7.2�?mol\\/L H2O2�67�?mol\\/L Fe(II), Au@Ag nanoparticles exhibited a resonance scattering (RS) peak at 538�nm. Upon addition of Catalase (Ct),\\u000a the
Aihui Liang; Yueyuan Liang; Zhiliang Jiang; Hesheng Jiang
Upregulation of intracellular antioxidant enzymes in brain and heart during estivation in the African lungfish Protopterus dolloi.
The African slender lungfish, Protopterus dolloi, is highly adapted to withstand periods of drought by secreting a mucous cocoon and estivating for periods of months to years. Estivation is similar to the diapause and hibernation of other animal species in that it is characterized by negligible activity and a profoundly depressed metabolic rate. As is typically observed in quiescent states, estivating P. dolloi are resistant to environmental stresses. We tested the hypothesis that P. dolloi enhances stress resistance during estivation by upregulating intracellular antioxidant defences in brain and heart tissues. We found that most of the major intracellular antioxidant enzymes, including the mitochondrial superoxide dismutase, cytosolic superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase, were upregulated in brain tissue of lungfish that had estivated for 60 days. Several of these enzymes were also elevated in heart tissue of estivators. These changes were not due to food deprivation, as they did not occur in a group of fish that were deprived of food but maintained in water for the same period of time. We found little evidence of tissue oxidative damage in estivators. Products of lipid peroxidation (4-hydroxynonenal adducts) and oxidative protein damage (carbonylation) were similar in estivating and control lungfish. However, protein nitrotyrosine levels were elevated in brain tissue of estivators. Taken together, these data indicate that estivating P. dolloi have enhanced oxidative stress resistance in brain and heart due to a significant upregulation of intracellular antioxidant capacity. PMID:19888582
Page, Melissa M; Salway, Kurtis D; Ip, Yuen Kwong; Chew, Shit F; Warren, Sarah A; Ballantyne, James S; Stuart, Jeffrey A
Peroxisomal Catalase in the Methylotrophic Yeast Candida boidinii: Transport Efficiency and Metabolic Significance
In this study we cloned CTA1, the gene encoding peroxisomal catalase, from the methylotrophic yeast Candida boidinii and studied targeting of the gene product, Cta1p, into peroxisomes by using green fluorescent protein (GFP) fusion proteins. A strain from which CTA1 was deleted (cta1? strain) showed marked growth inhibition when it was grown on the peroxisome-inducing carbon sources methanol, oleate, and d-alanine, indicating that peroxisomal catalase plays an important nonspecific role in peroxisomal metabolism. Cta1p carries a peroxisomal targeting signal type 1 (PTS1) motif, -NKF, in its carboxyl terminus. Using GFP fusion proteins, we found that (i) Cta1p is transported to peroxisomes via its PTS1 motif, -NKF; (ii) peroxisomal localization is necessary for Cta1p to function physiologically; and (iii) Cta1p is bimodally distributed between the cytosol and peroxisomes in methanol-grown cells but is localized exclusively in peroxisomes in oleate- and d-alanine-grown cells. In contrast, the fusion protein GFP-AKL (GFP fused to another typical PTS1 sequence, -AKL), in the context of CbPmp20 and d-amino acid oxidase, was found to localize exclusively in peroxisomes. A yeast two-hybrid system analysis suggested that the low transport efficiency of the -NKF sequence is due to a level of interaction between the -NKF sequence and the PTS1 receptor that is lower than the level of interaction with the AKL sequence. Furthermore, GFP-Cta1p?nkf coexpressed with Cta1p was successfully localized in peroxisomes, suggesting that the oligomer was formed prior to peroxisome import and that it is not necessary for all four subunits to possess a PTS motif. Since the main physiological function of catalase is degradation of H2O2, suboptimal efficiency of catalase import may confer an evolutionary advantage. We suggest that the PTS1 sequence, which is found in peroxisomal catalases, has evolved in such a way as to give a higher priority for peroxisomal transport to peroxisomal enzymes other than to catalases (e.g., oxidases), which require a higher level of peroxisomal transport efficiency.
Horiguchi, Hirofumi; Yurimoto, Hiroya; Goh, Toh-Kheng; Nakagawa, Tomoyuki; Kato, Nobuo; Sakai, Yasuyoshi
Immunodetection of a brown planthopper (Nilaparvata lugens St�l) salivary catalase-like protein into tissues of rice, Oryza sativa.
Saliva plays an important role in host plant-phloem-feeding insect molecular interactions. To better elucidate the role of insect saliva, a series of experiments were conducted to establish if catalase from the salivary glands of the brown planthopper (BPH; Nilaparvata lugens St�l) was secreted into rice host plant tissue during feeding. Catalase is the main enzyme that decomposes hydrogen peroxide (H2O2) at high concentrations. H2O2 is a part of the free radicals system that mediates important physiological roles including signalling and defence. Previous studies have suggested that H2O2 is involved in the rice endogenous response to BPH feeding. If, the BPH secretes catalase into host plant tissue this will counter the effects of H2O2, from detoxification to interfering with plant signalling and defence mechanisms. When BPHs were fed on a hopper-resistant rice variety for 24 h, catalase activity in the salivary glands increased 3.5-fold compared with hoppers fed on a susceptible rice variety. Further supporting evidence of the effects of BPH catalase was demonstrated by immunodetection analyses where results from two independent sources: BPH-infested rice tissue and BPH-probed artificial diets, suggest that the BPH secretes catalase-like protein during feeding. The possible physiological roles of BPH-secreted catalase are discussed. PMID:24164290
Petrova, A; Smith, C M
Screening of bacterial isolates from polluted soils exhibiting catalase and peroxidase activity and diversity of their responses to oxidative stress.
For the survival of individual isolates of gram-negative bacteria Pseudomonas putida, Achromobacter xylosoxidans, and the gram-positive bacterium Bacillus megaterium, in an environment polluted with crude oil products, the production of catalases exhibiting both catalase and dianisidine-peroxidase activity is important. Electrophoretic resolution of cell-free extracts of aerobically grown strains in Luria-Bertani medium during exponential phase revealed distinctive expression of catalatic and peroxidatic activities detected with 3,3'-diaminobenzidine tetrahydrochloride. A considerable diversity in microbial catalase and peroxidase responses to 20 or 40�mM H(2)O(2) stress, resulted from hydroperoxidase's variant of original isolates, indicating an environmental selective pressure. However, catalase was important for the adaptation of cultures to high concentration of 60�mM H(2)O(2). Appreciable differences in the sensitivity to toxic effect of H(2)O(2) (20 or 40�mM) treatment between individual isolates and their adapted variants during growth were observed until the middle of exponential phase, but they were insignificant at the entry to stationary phase. Isolates also exhibited a considerable diversity in catalases responses to phenolic contaminants 1 and 2�mM o- or p-phenylenediamine. Catalase activity of bacterium P.�putida was visibly stimulated only by p-phenylenediamine and not by its positional isomer o-PDA. This study contributes to a better understanding of the role catalases play in bacterial responses to a polluted environment. PMID:20145932
Buckov�, M�ria; Godoc�kov�, Jana; Z�mock�, Marcel; Polek, Bystr�k
The CYP2E1 inhibitor DDC up-regulates MMP-1 expression in hepatic stellate cells via an ERK1/2- and Akt-dependent mechanism
DDC (diethyldithiocarbamate) could block collagen synthesis in HSC (hepatic stellate cells) through the inhibition of ROS (reactive oxygen species) derived from hepatocyte CYP2E1 (cytochrome P450 2E1). However, the effect of DDC on MMP-1 (matrix metalloproteinase-1), which is the main collagen degrading matrix metalloproteinase, has not been reported. In co-culture experiments, we found that DDC significantly enhanced MMP-1 expression in human HSC (LX-2) that were cultured with hepatocyte C3A cells either expressing or not expressing CYP2E1. The levels of both proenzyme and active MMP-1 enzyme were up-regulated in LX-2 cells, accompanied by elevated enzyme activity of MMP-1 and decreased collagen I, in both LX-2 cells and the culture medium. H2O2 treatment abrogated DDC-induced MMP-1 up-regulation and collagen I decrease, while catalase treatment slightly up-regulated MMP-1 expression. These data suggested that the decrease in ROS by DDC was partially responsible for the MMP-1 up-regulation. ERK1/2 (extracellular signal-regulated kinase 1/2), Akt (protein kinase B) and p38 were significantly activated by DDC. The ERK1/2 inhibitor (U0126) and Akt inhibitor (T3830) abrogated the DDC-induced MMP-1 up-regulation. In addition, a p38 inhibitor (SB203580) improved MMP-1 up-regulation through the stimulation of ERK1/2. Our data indicate that DDC significantly up-regulates the expression of MMP-1�in LX-2 cells which results in greater MMP-1 enzyme activity and decreased collagen I. The enhancement of MMP-1 expression by DDC was associated with H2O2 inhibition and coordinated regulation by the ERK1/2 and Akt pathways. These data provide some new insights into treatment strategies for hepatic fibrosis.
Liu, Tianhui; Wang, Ping; Cong, Min; Xu, Youqing; Jia, Jidong; You, Hong
Beneficial effects of catalase or pyruvate in a most-probable-number technique for the detection of Staphylococcus aureus.
The effects of the addition of catalase (EC 220.127.116.11) or pyruvate on the enumeration of Staphylococcus aureus in Trypticase soy broth with 10% NaCl were examined using a most-probable-number technique. Addition of catalase or pyruvate to the broth increased enumeration of all heat-stressed S. aureus strains tested. Increases were also observed with nonstressed cells. Catalase and pyruvate were similarly effective when added to Trypticase soy broth-10% NaCl in enumerating staphylococci naturally present in low-temperature-rendered ground-beef samples.
Brewer, D G; Martin, S E; Ordal, Z J
Role of. pi. -cation radicals in the enzymatic cycles of peroxidases, catalases, and nitrite and sulfite reductases
Charge iterative extended Hueckel calculations, and magnetic and optical results on porphyrins, chlorins, and isobacteriochlorins (1) suggest that the catalytic cycles of the enzymes horseradish peroxidase, catalase, Neurospora crassa catalase, and nitrite and sulfite reductases proceed via ..pi..-cation radicals of their prosthetic groups; (2) offer distinguishing features for the optical and magnetic spectra of these radicals, pertinent to their detection as enzymatic intermediates; (3) reconcile the seemingly contradictory optical and NMR data on Compounds I of horseradish peroxidase; and (4) predict that the axial ligation of the heme differs for horseradish peroxidase and catalase.
Hanson, L K; Chang, C K; Davis, M S; Fajer, J
A Laboratory Experiment Investigating Different Aspects of Catalase Activity in an Inquiry - Based Approach
NASA Astrophysics Data System (ADS)
The action of the enzyme catalase on aqueous hydrogen peroxide to generate oxygen gas is a well-established demonstration (1-3). Catalase is typically obtained by aqueous extraction of a potato, and the potato extract is mixed together with 3% hydrogen peroxide. The oxygen that is produced can be collected over water. Variations on the procedure can demonstrate the dependence of catalytic activity on temperature or the presence of inhibitors (1, 2). The University of Colorado at Denver has used a version of this procedure as a laboratory in its second-semester course for nonmajors. Recently, students have been allowed to expand upon the procedures prescribed in the laboratory handout in an open-ended project format. We explored some of these variations in detail, and the results provided here offer ideas, centered around this laboratory, for open-ended projects that can be used in an inquiry-based approach.
Kimbrough, Doris R.; Magoun, Mary Ann; Langfur, Meg
Investigation of lipid peroxidation and catalase activity in magnetic fluid treated mice
NASA Astrophysics Data System (ADS)
The increasing interest in magnetic fluids (MFs) for biomedical applications demands a deeper knowledge of their effects in biological systems. To evaluate the in vivo response of a MF sample based on magnetite nanoparticles stabilized by a precoating double layer of dodecanoic acid plus ethoxylated polyalcohol (MFDE), the inflammation-related oxidative stress and antioxidant tissue response were both addressed in this study. MFDE sample was intraperitoneally administrated to mice at three different doses. The lipid peroxidation and the antioxidant defense induced in the liver and spleen were evaluated, respectively, by thiobarbituric acid-reactive substances (TBARS) and catalase activity, 1, 14, and 28 days after MFDE treatment. The liver and spleen responded with a huge increase in TBARS after MFDE treatment. We observed that oxidative changes as well as the variations in the liver catalase activity were time and MFDE-dose dependent.
Freitas, M. L. L.; Silva, L. P.; Freitas, J. L.; Azevedo, R. B.; Lacava, Z. G. M.; Homem de Bittencourt, P. I.; Curi, R.; Buske, N.; Morais, P. C.
Catalase is encoded by a multigene family in Arabidopsis thaliana (L.) Heynh.
The catalase multigene family in Arabidopsis includes three genes encoding individual subunits that associate to form at least six isozymes that are readily resolved by nondenaturing gel electrophoresis. CAT1 and CAT3 map to chromosome 1, and CAT2 maps to chromosome 4. The nucleotide sequences of the three coding regions are 70 to 72% identical. The amino acid sequences of the three catalase subunits are 75 to 84% identical and 87 to 94% similar, considering conservative substitutions. Both the individual isozymes and the individual subunit mRNAs show distinct patterns of spatial (organ-specific) expression. Six isozymes are detected in flowers and leaves and two are seen in roots. Similarly, mRNA abundance of the three genes varies among organs. All three mRNAs are highly expressed in bolts, and CAT2 and CAT3 are highly expressed in leaves.
Frugoli, J A; Zhong, H H; Nuccio, M L; McCourt, P; McPeek, M A; Thomas, T L; McClung, C R
Synthesis, cytotoxicity for mimics of catalase: Inhibitors of lactate dehydrogenase and hypoxia inducible factor.
Lactate dehydrogenase A (LDH-A) is a potentially important metabolic target for the inhibition of the highly activated glycolysis pathway in cancer cells. Two Mn(II) complexes with ligand containing di(pyridylmethyl) amine and pyrrol-ketone were used to attenuate the activity of LDH-A. The inhibition of the manganese(II) complexes on the proliferation of HepG-2 cells is related to their ability to disproportionate H2O2. Importantly, the synthesized mimic of catalase can decrease the expression of hypoxia inducible factor (HIF-1?) in HepG-2 cells. So we envision that the multifunctional mimics of catalase could attenuate the activity of LDH-A signaling the cancer cells to death through HIF-1? involved path. PMID:24763359
Xue, Juan-Juan; Chen, Qiu-Yun; Kong, Meng-Yun; Zhu, Chun-Yin; Gen, Zhi-Rong; Wang, Zhi-Lin
Cells contain a large number of antioxidants to prevent or repair the damage caused by ROS, as well as to regulate redox-sensitive signaling pathways General protocols are described to measure the antioxidant enzyme activity of superoxide dismutase (SOD), catalase, and glutathione peroxidase. The SODs convert superoxide radical into hydrogen peroxide and molecular oxygen, while the catalase and peroxidases convert hydrogen peroxide into water. In this way, two toxic species, superoxide radical and hydrogen peroxide, are converted to the harmless product water. Western blots, activity gels and activity assays are various methods used to determine protein and activity in both cells and tissue depending on the amount of protein needed for each assay. Other techniques including immunohistochemistry and immunogold can further evaluate the levels of the various antioxidant enzymes in tissue and cells. In general, these assays require 24 to 48 hours to complete.
Weydert, Christine J.; Cullen, Joseph J.
Enzymatic activity under tangential flow conditions of photochemically grafted membranes containing immobilized catalase.
Catalase has been immobilized within sandwich membranes prepared by the photoinduced grafting of an epoxy-diacrylate prepolymer onto commercial asymmetric cellulose membranes. The enzymatic activity of the membrane composite of hydrogen peroxide decomposition has been studied in a recirculation apparatus under tangential flow conditions without ultrafiltration. The enzymatic membranes were exposed to very low mechanical stresses and showed a very good catalytic performance and durability. Initial reaction rates, measured at 25 degrees C as a function of both substrate concentration and enzyme amount immobilized per unit membrane surface, indicate that the mechanism of action of catalase is not altered after immobilization, although substrate diffusion through the original thin layer of membranes may become rate controlling. PMID:18609576
Selli, E; D'Ambrosio, A; Bellobono, I R
Fibrin membrane endowed with biological function. V. Multienzyme complex of uricase, catalase, allantoinase and allantoicase.
The enzymes (uricase (EC 220.127.116.11), allantoinase (EC 184.108.40.206), and allantoicase (EC 18.104.22.168) which participate in degradation of purine bases, were embedded separately in fibrin membranes formed by fibrinogen-fibrin conversion with thrombin. All of these enzymes together with catalase were also embedded in a single fibrin membrane to make an immobilized multienzyme complex. The multienzyme complex in fibrin membrane thus prepared had an ability of degradation of uric acid to urea and glyoxylic acid via allantoin and allantoic acid. The stability of immobilized uricase or catalase embedded in fibrin membrane upon lyophilization was also tested in a comparison with nonimmobilized enzymes. PMID:6985799
Okamoto, H; Tipayang, P; Inada, Y
Catalase-Aminotriazole Method for Measuring Secretion of Hydrogen Peroxide by Microorganisms
A new method for measuring the secretion of H2O2 has been based upon an H2O2-dependent inhibition of catalase by 3-amino-1,2,4-triazole. The conversion of an H2O2-secretion rate into a catalase inhibition rate amplified a relatively small molar concentration of H2O2 and provided a highly specific and sensitive method for quantitatively measuring H2O2. A major advantage of this approach is that it does not require extensive accumulation of H2O2 in the environment. The method was successfully employed to measure H2O2 secretion by Mycoplasma pneumoniae, which possesses a peroxidase-like activity that limits the accumulation of H2O2 in the environment.
Cohen, Gerald; Somerson, Norman L.
Alterations in superoxide dismutase and catalase in Fusarium oxysporum during starvation-induced differentiation.
Vegetative hyphae of Fusarium oxysporum differentiate into chlamydospore by triggering with carbon-starvation. The current changes in the cellular detoxifying defenses against superoxide and hydrogen peroxide: superoxide dismutase (SOD) and catalase, were examined. Although there was a little change in catalase, a dramatic change in SOD was observed during the differentiation. In vegetative hyphae of F. oxysporum f. sp. raphani, three isozymes of SOD, all of which were not inhibited by hydrogen peroxide and cyanide, were present whereas in chlamydospore an isoenzyme, which was inhibited by hydrogen peroxide but not by cyanide, was present. Thus, as differentiation proceeded, Mn-type SOD disappeared and an Fe-type SOD appeared. The results suggest that the Fe-type SOD is specifically expressed during chlamydospore formation and that active intermediates of oxygen and/or its scavenging enzymes participate in the differentiation of Fusarium oxysporum. PMID:7626660
Kono, Y; Yamamoto, H; Takeuchi, M; Komada, H
Axenic growth up-regulates mass-specific metabolic rate, stress resistance, and extends life span in Caenorhabditis elegans.
Culture in axenic medium causes two-fold increases in the length of development and adult life span in Caenorhabditis elegans. We asked whether axenic medium imposes dietary restriction (ADR), and causes changes in metabolic activity and stress resistance. Eat mutants, which have a reduced food intake, were studied in parallel with wild-type worms to assess potential synergistic actions of axenic culture and food restriction. We found that axenic culture enhances metabolic activity as assessed by mass-specific oxygen consumption rate and heat production. Axenic culture also caused higher activities of the antioxidant enzymes superoxide dismutase and catalase, and led to increased resistance to high temperature, which was further exacerbated by mutation in eat-2. These results show that axenic medium up-regulates a variety of somatic maintenance functions including oxidative and thermal stress resistance and that food restriction due to axenic growth and to mutation in eat-2 are very similar but not identical. PMID:12559406
Houthoofd, Koen; Braeckman, Bart P; Lenaerts, Isabelle; Brys, Kristel; De Vreese, Annemie; Van Eygen, Sylvie; Vanfleteren, Jacques R
Isolation and characterization of two tightly linked catalase genes from castor bean that are differentially regulated
Two catalase genes,cat1 andcat2, have been isolated from the castor bean genome. They were located in the same direction on a chromosome at a distance of 2.4 kb,cat1 being on the downstream side ofcat2. The two genes contained introns at the same positions except that one of the 7 introns incat1 is missing incat2 and the corresponding introns differed in
Masaharu Suzuki; Takeshi Ario; Tsukaho Hattori; Kenzo Nakamura; Tadashi Asahi
Ergot cluster-encoded catalase is required for synthesis of chanoclavine-I in Aspergillus fumigatus
Genes required for ergot alkaloid biosynthesis are clustered in the genomes of several fungi. Several conserved ergot cluster\\u000a genes have been hypothesized, and in some cases demonstrated, to encode early steps of the pathway shared among fungi that\\u000a ultimately make different ergot alkaloid end products. The deduced amino acid sequence of one of these conserved genes (easC) indicates a catalase
Kerry E. Goetz; Christine M. Coyle; Johnathan Z. Cheng; Sarah E. O�Connor; Daniel G. Panaccione
Cytotoxicity of lawsone and cytoprotective activity of antioxidants in catalase mutant Escherichia coli.
Lawsone is an active naphthoquinone derivative isolated from henna (Lawsonia inermis L.), a widely used hair dye. Previous study on the toxicity of lawsone remains unclear since the involvement of oxidative stress and the kind of ROS (reactive oxygen species) involved have not been fully resolved yet. This present study reports the cytotoxic effects of lawsone and henna. We carried out CAT assay (a zone of inhibition test of bacterial growth and colony-forming efficiency test of transformant Escherichia coli strains that express mammalian catalase gene derived from normal catalase mice (Cs(a)) and catalase-deficient mutant mice (Cs(b))), Ames mutagenicity assay and H(2)O(2) generation assay. Lawsone generated H(2)O(2) slightly in phosphate buffer system and was not mutagenic in Ames assay using TA 98, TA 100 and TA 102, both in the absence and presence of metabolic activation. Lawsone exposure inhibited the growth of both Cs(a) and Cs(b) strains in a dose-dependent manner. Mean zone diameter for Cs(a) was 9.75+/-0.96 mm and 12.75+/-1.5 mm for Cs(b). Natural henna leaves did not show toxic effects, whereas two out of four samples of marketed henna products were shown toxicity effects. Catalase abolished zone of inhibition (ZOI) of marketed henna products, eliminated ZOI of lawsone in a dose-dependent manner and low concentration of exogenous MnSOD and Cu/ZnSOD eliminated the toxicity. Histidine and DTPA, the metal chelator; BHA and low concentration of capsaicin, the inducer of NADH-quinone reductase, effectively protected Cs(a) and Cs(b) against lawsone in this study. We suggest that lawsone cytotoxicity is probably mediated, at least in part, by the release of O(2)(-), H(2)O(2) and OH(-). PMID:17442476
Sauriasari, Rani; Wang, Da-Hong; Takemura, Yoko; Tsutsui, Ken; Masuoka, Noriyoshi; Sano, Kuniaki; Horita, Masako; Wang, Bing-Ling; Ogino, Keiki
Adenovirus-Mediated Gene Transfer of Superoxide Dismutase and Catalase Decreases Restenosis after Balloon Angioplasty
Background: Reactive oxygen species (ROS) production increases after injury and potentially contributes to restenosis after angioplasty. We therefore evaluated the effect of adenovirus-mediated gene transfer (Ad) of superoxide dismutase (SOD) and catalase (CAT) on ROS production and restenosis after balloon angioplasty. Methods: O2� and H2O2 production was quantified in cultured cells after incubation with either LPS or CuSO4. Angioplasty and
Eric Durand; Ayman Al Haj Zen; Faouzi Addad; Camille Brasselet; Giuseppina Caligiuri; Fran�ois Vinchon; Patricia Lemarchand; Michel Desnos; Patrick Bruneval; Antoine Lafont
Angiotensin-(1-7) prevents diabetes-induced attenuation in PPAR-? and catalase activities
The mechanisms by which angiotensin-(1�7) [Ang-(1�7)] exerts its beneficial effects on end-organ damage associated with diabetes and hypertension are not well understood. The purpose of this study was A) to compare the effects of apocynin with Ang-(1�7) on renal vascular dysfunction and NADPH oxidase activity in a combined model of diabetes and hypertension and B) to further determine whether chronic treatment with Ang-(1�7) can modulate renal catalase, and peroxisome proliferator activated receptor-? (PPAR�?) levels in streptozotocin induced-diabetes in both normotensive Wistar Kyoto rats (WKY) and in spontaneously hypertensive rats (SHR). Apocynin or Ang-(1�7) treatment for one month starting at the onset of diabetes similarly attenuated elevation of renal NADPH oxidase activity in the diabetic SHR kidney and reduced the degree of proteinuria and hyperglycemia, but had little or modest effect on reducing mean arterial pressure. Both drugs also attenuated the diabetes-induced increase in renal vascular responsiveness to endothelin-1. Induction of diabetes in WKY and SHR animals resulted in significantly reduced renal catalase activity and in PPAR�? mRNA and protein levels. Treatment with Ang-(1�7) significantly prevented diabetes-induced reduction in catalase activity and the reduction in PPAR�? mRNA and protein levels in both animal models. Taken together, these data suggest that activation of Ang-(1�7)-mediated signaling could be an effective way to prevent the elevation of NADPH oxidase activity and inhibition of PPAR�? and catalase activities in diabetes and/or hypertension.
Dhaunsi, Gursev S.; Yousif, Mariam H. M.; Akhtar, Saghir; Chappell, Mark C.; Diz, Debra I.; Benter, Ibrahim F.
Post-Transcriptional Regulation of Catalase Isozyme Expression in Cotton Seeds.
We reported previously that expression of the five tetrameric catalase isozymes during postgerminative growth of cotton seedings was a consequence of interactions between two subunits (SU 1 and SU 2) temporally synthesized from two distinct catalase genes. In this study, we focused on the regulation of the expression of these two catalase subunits during the changeover from glyoxysomal to leaf-type peroxisomal metabolism. The steady-state level of glyoxysomal SU 1 protein (present in 12-hour-old seeds) increased through day 3 and then declined linearly through day 6, whereas SU 2 protein (first detected in 24-hour-old seeds) increased continuously through day 6. The time courses for steady-state levels of the mRNAs encoding these two subunits revealed two clearly separated peaks: the first at day 1 (SU 1) and the other at day 4 (SU 2). Accumulation of these mRNAs preceded the accumulation of their corresponding proteins by at least 24 hours, suggesting temporal, pretranslational regulation of synthesis of both subunits. Results from run-on transcriptional assays with isolated nuclei, however, revealed that transcripts encoding both subunits were synthesized together on days 1 through 5. Hence, it appears that the accumulations of SU 1 and SU 2 mRNAs are controlled primarily at the post-transcriptional level, which has not been reported for catalase or any other eukaryotic peroxisomal enzymes. The accumulation of SU 1 mRNA is not light dependent, whereas the accumulation of SU 2 mRNA, which directs synthesis of the predominant subunit comprising the leaf-type peroxisomal isozyme, occurs only after exposure of seedlings to light.
Ni, W; Trelease, RN
Murine and human b locus pigmentation genes encode a glycoprotein (gp75) with catalase activity
Melanogenesis is regulated in large part by tyrosinase, and defective tyrosinase leads to albinism. The mechanisms for other pigmentation determinants (e.g., those operative in tyrosinase-positive albinism and in murine coat-color mutants) are not yet known. One murine pigmentation gene, the brown (b) locus, when mutated leads to a brown (b/b) or hypopigmentated (B{sup lt}/B{sup lt}) coat versus the wild-type black (B/B). The authors show that the b locus codes for a glycoprotein with the activity of a catalase (catalase B). Only the c locus protein is a tyrosinase. Because peroxides may be by-products of melanogenic activity and hydrogen peroxide in particular is known to destroy melanin precursors and melanin, they conclude that pigmentation is controlled not only by tyrosinase but also by a hydroperoxidase. The studies indicate that catalase B is identical with gp75, a known human melanosomal glycoprotein; that the b mutation is in a heme-associated domain; and that the B{sup lt} mutation renders the protein susceptible to rapid proteolytic degradation.
Halaban, R.; Moellmann, G. (Yale Univ. School of Medicine, New Haven, CT (USA))
Cytoplasmic superoxide dismutase and catalase activity and resistance to radiation lethality in murine tumor cells
Reduced species of molecular oxygen are produced by the interaction of ionizing radiation with aqueous solutions containing molecular oxygen. The enzymes catalase and superoxide dismutase (SOD) are thought to function in vivo as scavengers of metabolically produced peroxide and superoxide respectively. SOD has been shown to protect against the lethal effects of ionizing radiation in vitro and in vivo. The authors have investigated the relationship between the cytosolic SOD catalase content and the sensitivity to radiation lethality of a number of murine cell lines (402AX, EL-4, MB-2T3, MB-4, MEL, P-815, SAI, SP-2, and SV-3T3). K/sub i/(CN/sup -/) for murine Cu-Zn-SOD was determined to be 6.8 x 10/sup -6/ M. No cytosolic Mn-SOD activity was found in any of the cell lines studied. No correlation was found between the cytosolic Cu-Zn-SOD or cytosolic catalase activity and the resistance to radiation lethality or the murine cell lines studied.
Davy, C.A.; Tesfay, Z.; Jones, J.; Rosenberg, R.C.; McCarthy, C.; Rosenberg, S.O.
How catalase recognizes H?O? in a sea of water.
Monofunctional heme-catalases have been studied for many decades but there is still an incomplete understanding of why such a large tetrameric protein with deeply buried active sites is required to accomplish such a simple reaction as H2 O2 dismutation. Catalase accomplishes this reaction at a high rate although water at 55 M is expected to compete with H2 O2 for the enzyme's active site. Using molecular dynamics simulations we addressed the question as to how catalase selects H2 O2 in water. Selection is accomplished through different mechanisms: higher residence time of H2 O2 in the vicinity of certain prevalent amino acid residues at the protein surface and substrate channel, coordinated motion of the main passage amino acids that is increased in the presence of H2 O2 , a gate valve mechanism consisting of the motion of two contiguous phenylalanine residues that drive water molecules out of the final section of the substrate channel, a hydrophobic barrier before the active site that was crossed more easily by H2 O2 which kept most of its hydrogen bonds while passing, and finally an increased residence time for H2 O2 at the active site. These mechanisms, based on the physicochemical differences between H2 O2 and water, provide an explanation as to why such a large tetrameric protein with deeply buried active sites is required to accomplish efficient H2 O2 dismutation. PMID:23818262
Dom�nguez, Laura; Sosa-Peinado, Alejandro; Hansberg, Wilhelm
Prevalence of Catalase (-21 A/T) Gene Variant in South Indian (Tamil) Population
Catalase, an endogenous antioxidant enzyme, is responsible for regulating reactive species levels. Several epidemiologic studies have suggested that single nucleotide polymorphism in catalase gene may be associated with many diseases. The genotype of CAT (-21 A/T) point mutation in promoter region of catalase gene was determined by polymerase chain based restriction fragment length polymorphism analysis in the DNA of 100 healthy volunteers. The frequency of CAT (-21 A/T) gene polymorphism AA, AT, and TT genotypes was found to be 7, 23, and 70 percent, respectively. The mutant �T� allele frequency was found to be 0.82 among the south Indian (Tamil) population. Chi square analysis showed that the study population lies within the Hardy-Weinberg equilibrium. The wild type genotype (AA) was found to be very low (7%) and the mutant genotype (AT/TT) was found to be more prevalent (93%) among the south Indian population. This suggests that the high prevalence of mutant genotype may increase the susceptibility to oxidative stress associated diseases.
Lourdhu Mary, A.; Nithya, K.; Isabel, W.
Intracellular targeting of ascomycetous catalase-peroxidases (KatG1s).
Bifunctional catalase-peroxidases (KatGs) are heme oxidoreductases widely spread among bacteria, archaea and among lower eukaryotes. In fungi, two KatG groups with different localization have evolved, intracellular (KatG1) and extracellular (KatG2) proteins. Here, the cloning, expression analysis and subcellular localization of two novel katG1 genes from the soil fungi Chaetomium globosum and Chaetomium cochliodes are reported. Whereas, the metalloenzyme from Ch. globosum is expressed constitutively, Ch. cochliodes KatG1 reveals a slight increase in expression after induction of oxidative stress by cadmium ions and hydrogen peroxide. The intronless open reading frames of both Sordariomycetes katG1 genes as well as of almost all fungal katG1s possess two peroxisomal targeting signals (PTS1 and PTS2). Peroxisomal localization of intracellular eukaryotic catalase-peroxidases was verified by organelle separation and immunofluorescence microscopy. Co-localization with the peroxisomal enzyme 3-ketoacyl-CoA-thiolase was demonstrated for KatGs from Magnaporthe grisea, Chaetomium globosum and Chaetomium cochliodes. The physiological role of fungal catalase-peroxidases is discussed. PMID:23589225
Z�mock�, Marcel; Sekot, Gerhard; Bu?kov�, M�ria; Godo?�kov�, Jana; Sch�ffer, Christina; Farka�ovsk�, Mari�n; Obinger, Christian; Polek, Bystr�k
Contribution of superoxide dismutase and catalase activities to Shigella flexneri pathogenesis.
A Shigella flexneri serotype 5 strain deficient in the production of the iron-containing superoxide dismutase FeSOD (sodB) and a catalase-negative (katFG) S. flexneri serotype 5 strain were isolated. Both strains were examined for increased sensitivity to oxygen stress by using assays involving killing by mouse peritoneal macrophages and human polymorphonuclear leukocytes as well as infection of rabbit ileal loops. The sodB mutant was extremely sensitive to killing by phagocytes when compared with the wild-type parent, M90T. The catalase mutant also showed an increased sensitivity to killing, but to a much lesser extent. Upon infection of rabbit ileal loops and subsequent histopathological examination, the sodB mutant caused very little detectable damage to intestinal villi. The pattern of infection was roughly similar to that of BS176, an avirulent plasmidless derivative of M90T. The katFG mutant, on the other hand, showed a high degree of destruction, similar to that caused by M90T. This evidence suggests that the superoxide dismutase encoded by sodB may play an important role in the pathogenesis of S. flexneri. In contrast, catalases appear to make a limited contribution to virulence. Images
Franzon, V L; Arondel, J; Sansonetti, P J
Fe(III) complexes of 1,4,8,11-tetraaza[14]annulenes as catalase mimics.
The development of enzyme mimics of catalase which decompose hydrogen peroxide to water and molecular oxygen according to the 2:1 stoichiometry of native catalase and in aqueous solution at pH 7 and at micromolar concentrations of the enzyme model and hydrogen peroxide is reported. For this purpose, iron(III) complexes of 1,4,8,11-tetraaza[14]annulenes are prepared by various procedures. Efficacious preparations utilize reaction of the [N4] macrocyles with FeII salts in the presence of triphenylamine, followed by gentle oxidation of the FeII complexes by molecular oxygen or by tris(4-bromophenyl)aminium hexachloroantimonate. The complexes are characterized by SQUID magnetometry and by M�ssbauer, EPR, and UV/vis spectrometry. In the solid state, the iron(III) center of the catalytically active complexes exists in the intermediate (quartet, S = 3/2) spin state. Several of these complexes decompose hydrogen peroxide in aqueous buffer solution at pH 7.2 at room temperature with turnover numbers between 40 and 80. The apparent second-order rate constant for hydrogen peroxide decomposition is in the range of 1400-2400 M(-1) s(-1), about 3 orders of magnitude lower than the value for native catalase. Besides oxygen production, a non-oxygen releasing pathway of hydrogen peroxide decomposition is unveiled. PMID:18001111
Sustmann, Reiner; Korth, Hans-Gert; Kobus, Diana; Baute, J�rg; Seiffert, Karl-Heinz; Verheggen, Elisabeth; Bill, Eckhard; Kirsch, Michael; de Groot, Herbert
Terazosin-induced alterations in catalase expression and lipid peroxidation in the rat seminal vesicles.
Previous studies have shown that alpha1-adrenergic receptor antagonists may alter seminal vesicle contractility and impair fertility in male rats. This study was designed to investigate the effects of terazosin on the catalase expression in the seminal vesicles and the lipid peroxidation of the seminal fluid in normal adult rats. Wistar rats were treated with terazosin (1.2 mg kg(-1) body weight, given orally every second day) for 120 days. Catalase expression was assessed immunohistochemically in tissue sections of the seminal vesicles, and lipid peroxidation was estimated by measuring the malondialdehyde (MDA) levels in the seminal vesicles' fluid. The seminal vesicles in terazosin-treated rats were particularly distended in comparison with those of controls, and their secreting epithelium was suppressed. Cytoplasmic catalase expression in the secreting epithelial cells (% of cells) was increased in terazosin-treated specimens in comparison with controls (76.1 � 17.1 versus 51.3 � 25.1, P = 0.005). MDA levels (?m) were also higher in samples from treated subjects in comparison with controls (2.67 � 1.19 versus 1.39 � 0.19, P = 0.01). Although the direct effect of terazosin treatment on the seminal vesicles is that of impaired contractility, an indirect effect is that on fertility by increasing lipid peroxidation in the seminal fluid and/or through degrading of hydrogen peroxide that is essential for sperm capacitation. PMID:22731390
Mitropoulos, D; Patris, E; Deliconstantinos, G; Kyroudi-Voulgari, A; Anastasiou, I; Perea, D
Inactivation of catalase by phenylhydrazine. Formation of a stable aryl-iron heme complex.
Catalase promotes the H2O2-dependent oxidation of phenylhydrazine to benzene but simultaneously is subject to a pseudo-first order inactivation process. Each inactivation event is subtended by catalytic turnover of three molecules of phenylhydrazine and 52 molecules of H2O2. The dimethyl ester of N-phenylprotoporphyrin IX is extracted with acidic methanol from the inactivated enzyme, but the prosthetic heme with a phenyl sigma-bonded to the iron atom is obtained by gentle extraction with 2-butanone. The absolute chirality of N-ethylprotoporphyrin IX isolated from catalase inactivated with ethylhydrazine confirms that the prosthetic heme has the same chiral orientation in the active site as it does in hemoglobin. The known inactivation of methemoglobin by phenylhydrazine is shown to depend on H2O2 but not oxygen. The results demonstrate that the H2O2-dependent oxidation of phenylhydrazine by catalase and other hemoproteins results in sigma-coordination of a phenyl residue to the prosthetic heme iron. This process may play a role not only in phenylhydrazine-mediated erythrocyte lysis but also in the activation of guanylate cyclase. PMID:6885792
Ortiz de Montellano, P R; Kerr, D E
Reactive oxygen species up-regulate CD11b in microglia via nitric oxide: Implications for neurodegenerative diseases
Microglial activation is considered as a hallmark of several neurodegenerative disorders. During microglial activation, the expression of CD11b, the beta-integrin marker of microglia, is increased. However, the molecular mechanism behind increased microglial CD11b expression is poorly understood. The present study was undertaken to explore the role of reactive oxygen species (ROS) in the expression of CD11b in microglial cells. Bacterial lipopolysaccharide (LPS) stimulated the expression of CD11b in mouse BV-2 microglial cells and primary microglia, the effect that was blocked by antioxidants such as N-acetylcysteine (NAC) and pyrrolidine dithiocarbamate (PDTC). Furthermore, comicroinjection of either NAC or PDTC with LPS was also able to suppress LPS-stimulated expression of CD11b in striatum in vivo. Similarly, other neurotoxic molecules, such as interleukin-1? (IL-1?), IL-12 p402, fibrillar amyloid-? (A?) peptides, HIV-1 gp120, and double-stranded RNA (poly(IC)), also stimulated the expression of CD11b in microglia through the involvement of ROS. Complete inhibition of LPS-stimulated expression of CD11b by catalase, induction of CD11b expression by H2O2 alone, and inhibition of superoxide-stimulated CD11b expression by catalase suggest that H2O2, but not superoxide, is in fact involved in the expression of CD11b. Interestingly, we also demonstrate that ROS stimulated the expression of CD11b after the induction of nitric oxide (NO) production and failed to stimulate CD11b when NO production was inhibited by either 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (carboxy-PTIO) or L-N6-(1-iminoethyl)-L-lysine (L-NIL). Taken together, these studies suggest that the up-regulation of CD11b in microglia is redox sensitive and that ROS up-regulates CD11b via NO.
Roy, Avik; Jana, Arundhati; Yatish, Kavitha; Freidt, Matthew B.; Fung, Yiu K.; Martinson, Jeffrey A.; Pahan, Kalipada
Using superoxide dismutase/catalase mimetics to manipulate the redox environment of neural precursor cells.
Past work has shown that neural precursor cells are predisposed to redox sensitive changes, and that oxidative stress plays a critical role in the acute and persistent changes that occur within the irradiated CNS. Irradiation leads to a marked rise in reactive oxygen species (ROS) that correlates with oxidative endpoints in vivo and reductions in neurogenesis. To better understand the impact of oxidative stress on neural precursor cells, and to determine if radiation-induced oxidative damage and precursor cell loss after irradiation could be reduced, a series of antioxidant compounds (EUK-134, EUK-163, EUK-172, EUK-189) were tested, three of which possess both superoxide dismutase (SOD) and catalase activities and one (EUK-163) whose only significant activity is SOD. Our results show that these SOD/catalase mimetics apparently increase the oxidation of a ROS-sensitive fluorescent indicator dye, particularly after short (12 h) treatments, but that longer treatments (24 h) decrease oxidation attributable to radiation-induced ROS. Similarly, other studies found that cells incubated with CuZnSOD showed some increase in intracellular ROS levels. Subsequent data suggested that the dye-oxidising capabilities of the EUK compounds were linked to differences in their catalase activity and, most likely, their ability to catalyse peroxidative pathways. In unirradiated mice, the EUK-134 analogue induced some decrease of proliferating precursor cells and immature neurons 48 h after radiation, an effect that may be attributable to cytotoxicity and/or inhibition of precursor proliferation. In irradiated mice, a single injection of EUK-134 was not found to be an effective radioprotector at acute times (48 h). The present results support continued development of our in vitro model as a tool for predicting certain in vivo responses, and suggest that in some biological systems the capability to scavenge superoxide but produce excess H(2)O(2), as is known for CuZnSOD, may be potentially deleterious. Our results also show that the ability of catalase mimetics, like true catalases, to catalyse peroxidase reactions can complicate the interpretation of data obtained with certain fluorescent ROS-indicator dyes. PMID:17166877
Limoli, C L; Giedzinski, E; Baure, J; Doctrow, S R; Rola, R; Fike, J R
Evaluation of criteria used in the identification of actinomyces bovis with particular reference to the catalase reaction
The identification of smooth typeActinomyces bovis is an extremely difficult one to make. By a well selected set of criteria, including the catalase test, it is generally possible to make an unquestionable identification.
Loyal S. Suter
Effects of the seminal plasma zinc content and catalase activity on the semen quality of water buffalo (Bubalus bubalis) bulls.
In order to determine zinc and catalase content of seminal plasma in the buffalo and to study their associations with the semen characteristics, 54 semen samples were collected from 10 buffalo bulls; semen volume and sperm concentration, gross and progressive motility and viability were evaluated, seminal plasma was then harvested by centrifugation and its zinc content was estimated by atomic absorption spectrophotometer and its catalase activity determined by using a commercial kit. The zinc content of the seminal plasma (Mean +/- SEM) was recorded as 154.40 +/- 1.74 mg L(-1), while, the mean catalase value was 32.00 +/- 0.42 U mL(-1). The mean zinc values was highly correlated with sperm progressive motility and viability and with catalase values (p = 0.000 for all) and also was associated with gross motility (p = 0.020) and negatively with abnormal morphology (p = 0.049). The catalase values were highly associated with sperm progressive motility, viability and zinc content (p = 0.000 for all) and was associated with sperm gross motility (p = 0.024). For further clarification of these correlations, the samples were categorized in three groups of excellent (Ex, >90% motile, n = 33), good (Go, 80-89% motile, n = 15) and moderate (Mo, <79% motile, n = 6) according to their percentage of sperm motility. The mean progressive motility in Ex group was 92.54 +/- 0.51%, in Go group was 81.66 +/- 0.62% and in Mo group was 71.66 +/- 1.05%. The mean zinc and catalase values were recorded as 161.07 +/- 1.63 mg L(-1) and 33.41 +/- 0.34 U mL(-1) in Ex, 146.70 +/- 1.91 mg L(-1) and 31.01 +/- 0.67 in Go and 136.42 +/- 4.97 mg L(-1) and 26.51 +/- 0.87 U mL(-1) in Mo groups. The mean zinc value in Ex group was highly associated with sperm motility, viability and catalase values, in Go group was associated with catalase values and highly associated with sperm abnormal morphology and in Mo group it was highly associations with catalase values only. The mean catalase value in Ex group, was highly associated with sperm motility and viability, in Go group was associated with zinc content and in Mo groups was highly associated with the zinc content. These results show that seminal plasma zinc and catalase content are correlated with semen characteristics and synergistically act to preserve motility and viability of the spermatozoa after ejaculation. PMID:19579933
Alavi-Shoushtari, S M; Rezai, S Asri; Ansari, M H Kh; Khaki, A
Catalase and alternative oxidase cooperatively regulate programmed cell death induced by ?-glucan elicitor in potato suspension cultures
In potato (Solanum tuberosum L.) suspension cells, the expression of the gene encoding alternative oxidase (AOX) and H2O2 accumulation were induced by treatment with �-glucan elicitor. The inhibition of catalase activity enhanced both AOX mRNA expression and the production of H2O2, whereas the ascorbate peroxidase inhibitor did not have any effect on these responses. Simultaneous inhibition of catalase and AOX
Masashi Mizuno; Yasuomi Tada; Kimitaka Uchii; Sachiko Kawakami; Shigeyuki Mayama
An immobilised catalase peroxidase from the alkalothermophilic Bacillus SF for the treatment of textile-bleaching effluents
A catalase peroxidase (CP) from the newly isolated Bacillus SF was used to treat textile-bleaching effluents. The enzyme was stable at high pH values and temperatures, but was more sensitive to deactivation by hydrogen peroxide than monofunctional catalases. Based on the Michaelis-Menten kinetics of the CP, a model was developed to describe its deactivation characteristics. The enzyme was immobilised on
G. Fruhwirth; A. Paar; M. Gudelj; Austria A. Cavaco-Paulo; K.-H. Robra; G. G�bitz
Active and inhibited human catalase structures: ligand and NADPH binding and catalytic mechanism 1 1 Edited by R. Huber
Human catalase is an heme-containing peroxisomal enzyme that breaks down hydrogen peroxide to water and oxygen; it is implicated in ethanol metabolism, inflammation, apoptosis, aging and cancer. The 1.5 � resolution human enzyme structure, both with and without bound NADPH, establishes the conserved features of mammalian catalase fold and assembly, implicates Tyr370 as the tyrosine radical, suggests the structural basis
Christopher D Putnam; Andrew S Arvai; Yves Bourne; John A Tainer
Characterization of transgenic tomato plants expressing an antisense catalase gene and cloning of a TOMCAT2 gene
Transgenic tomatoes (Lycopersicon esculentum Mill. �Ohio 8245�) expressing an antisense catalase gene (ASTOMCAT1) were used to test the hypothesis that modification of the reactive oxygen species scavenging mechanism in plants can lead to changes in oxidative strew tolerance. A two to eight-fold reduction in total catalase activity and a two-fold increase in levels of H2O2 were detected in the leaf
Kanogwan Kerdnaimongkol
Cardiac-specific overexpression of catalase prolongs lifespan and attenuates ageing-induced cardiomyocyte contractile dysfunction and protein damage.
1. Oxidative stress plays a role in senescence-associated organ deterioration. This is supported by the beneficial effects of anti-oxidants against ageing-related organ damage, although their role in cardiac ageing has not been elucidated. 2. The aim of the present study was to examine the impact of cardiac-specific overexpression of catalase, an enzyme for H(2)O(2) detoxification, on cardiac contractile function and protein damage in young (3-4 months) and old (26-28 months) male mice. Lifespan was analysed using the Kaplan-Meier survival curve. Cardiomyocyte contractile indices at various stimulus frequencies (0.1-5.0 Hz) were analysed, including peak shortening (PS), time to 90% PS, time to 90% relengthening (TR(90)) and maximal velocity of shortening/relengthening (+/-dL/dt). Protein damage was assessed using protein carbonyl formation. Catalase transgenic mice showed longer lifespan than wild-type FVB mice. The catalase transgene itself did not alter bodyweight or organ weight, or myocyte function. Ageing depressed +/-dL/dt and prolonged TR(90), but had no effect on other indices in FVB mice. Increased frequency triggered decreases in PS amplitude were exaggerated in aged FVB myocytes. Interestingly, ageing-induced mechanical defects were significantly attenuated in myocytes from catalase mice. Protein carbonyl formation was elevated in aged FVB compared with young FVB mice, which was significantly diminished in catalase mice. The proteomes of the myocardium of young or old FVB and catalase mice were compared using two-dimensional gel electrophoresis and mass spectrometry. Six proteins with differential expression between young and old FVB groups were tentatively identified, some of which were reversed by catalase. 3. In summary, the present data suggest that catalase protects cardiomyocytes from ageing-induced contractile defects and protein damage. PMID:17201740
Wu, Shan; Li, Qun; Du, Min; Li, Shi-Yan; Ren, Jun
Catalase activity in macro- and microorganisms as an indicator of biotic stress in coastal waters of the eastern Mediterranean Sea
In this study we examined the activity of catalase in the water column (mainly attributed to planktonic microorganisms) and\\u000a the activity of catalase and superoxide dismutase (SOD), as well as lipid peroxidation in the midgut gland of the benthic\\u000a bivalve Donax trunculus as possible indicators of biotic stress. The measurements were performed at stations situated at known contaminated and clean
D. L. Angel; U. Fiedler; N. Eden; N. Kress; D. Adelung; B. Herut
Catalase potentiates retinoic acid-induced THP1 monocyte differentiation into macrophage through inhibition of peroxisome proliferator-activated receptor
Macrophage differentiation plays a piv- otal role in cardiovascular diseases and many other physiological processes. However, the role of re- action oxygen species in macrophage differentia- tion has not been elucidated. Here, we report func- tional characterization of catalase, an enzyme that degrades hydrogen peroxide (H2O2), in THP-1 monocyte differentiation. Treatment of THP-1 cells with catalase was able to synergize
Qiurong Ding; Ting Jin; Zhenzhen Wang; Yan Chen
Expression, purification, and sequence analysis of catalase-1 from the soil bacterium Comamonas terrigena N3H
Catalases are essential components of the cellular equipment to cope with oxidative stress. We have purified and characterize herein the most abundant heme-containing catalase-1 from the soil bacterium Comamonas terrigena N3H. This oxidative stress-induced enzyme was isolated from exponential phase cells grown in the presence of peroxyacetic acid. We have used consecutive steps of hydrophobic, molecular sieve, and ion exchange
Marcel Z�mock�; Jana Godo???kov�; Juraj Ga�per??k; Franz Koller; Bystr??k Polek
The effect of oxidative stress on the mutation rate of Mycobacterium tuberculosis with impaired catalase\\/peroxidase function
Objectives: To determine the effect of oxidative stress on isoniazid-resistant Mycobacterium tuber- culosis deficient in catalase\\/peroxidase activity to varying degrees through mutation in katG. Methods: The mutation rate was determined for a set of isogenic strains with different katG alleles giving different catalase and\\/or peroxidase activities following exposure to the oxidizing agent, hydro- gen peroxide. Mutants were selected on rifampicin,
D. M. O'Sullivan; T. D. McHugh; S. H. Gillespie
Direct electrochemistry and electrocatalytic activity of catalase immobilized onto electrodeposited nano-scale islands of nickel oxide.
Cyclic voltammetry was used for simultaneous formation and immobilization of nickel oxide nano-scale islands and catalase on glassy carbon electrode. Electrodeposited nickel oxide may be a promising material for enzyme immobilization owing to its high biocompatibility and large surface. The catalase films assembled on nickel oxide exhibited a pair of well defined, stable and nearly reversible CV peaks at about -0.05 V vs. SCE at pH 7, characteristic of the heme Fe (III)/Fe (II) redox couple. The formal potential of catalase in nickel oxide film were linearly varied in the range 1-12 with slope of 58.426 mV/pH, indicating that the electron transfer is accompanied by single proton transportation. The electron transfer between catalase and electrode surface, (k(s)) of 3.7(+/-0.1) s(-1) was greatly facilitated in the microenvironment of nickel oxide film. The electrocatalytic reduction of hydrogen peroxide at glassy carbon electrode modified with nickel oxide nano-scale islands and catalase enzyme has been studied. The embedded catalase in NiO nanoparticles showed excellent electrocatalytic activity toward hydrogen peroxide reduction. Also the modified rotating disk electrode shows good analytical performance for amperometric determination of hydrogen peroxide. The resultant catalase/nickel oxide modified glassy carbon electrodes exhibited fast amperometric response (within 2 s) to hydrogen peroxide reduction (with a linear range from 1 microM to 1 mM), excellent stability, long term life and good reproducibility. The apparent Michaelis-Menten constant is calculated to be 0.96(+/-0.05)mM, which shows a large catalytic activity of catalase in the nickel oxide film toward hydrogen peroxide. The excellent electrochemical reversibility of redox couple, high stability, technical simplicity, lake of need for mediators and short preparations times are advantages of this electrode. Finally the activity of biosensor for nitrite reduction was also investigated. PMID:17166647
Salimi, Abdollah; Sharifi, Ensiyeh; Noorbakhsh, Abdollah; Soltanian, Saied
Layer-by-layer immobilized catalase on electrospun nanofibrous mats protects against oxidative stress induced by hydrogen peroxide.
Catalase, a kind of redox enzyme and generally recognized as an efficient agent for protecting cells against hydrogen peroxide (H2O2)-induced cytotoxicity. The immobilization of catalase was accomplished by depositing the positively charged chitosan and the negatively charged catalase on electrospun cellulose nanofibrous mats through electrospining and layer-by-layer (LBL) techniques. The morphology obtained from Field emission scanning electron microscopy (FE-SEM) indicated that more orderly arranged three-dimension (3D) structure and roughness formed with increasing the number of coating bilayers. Besides, the enzyme-immobilized nanofibrous mats were found with high enzyme loading and activity, moreover, X-ray photoelectron spectroscopy (XPS) results further demonstrated the successful immobilization of chitosan and catalase on cellulose nanofibers support. Furthermore, we evaluated the cytotoxicity induced by hydrogen peroxide in the Human umbilical vascular endothelial cells with or without pretreatment of nanofibrous mats by MTT assay, LDH activity and Flow cytometric evaluation, and confirmed the pronounced hydrogen peroxide-induced toxicity, but pretreatment of immobilized catalase reduced the cytotoxicity and protected cells against hydrogen peroxide-induced cytotoxic effects which were further demonstrated by scanning electron microscopy (SEM) and Transmission Electron Microscopy (TEM) images. The data pointed toward a role of catalase-immobilized nanofibrous mats in protecting cells against hydrogen peroxide-induced cellular damage and their potential application in biomedical field. PMID:24804555
Huang, Rong; Deng, Hongbing; Cai, Tongjian; Zhan, Yingfei; Wang, Xiankai; Chen, Xuanxuan; Ji, Ailing; Lil, Xueyong
Expression and purification of soluble bio-active rice plant catalase-A from recombinant Escherichia coli.
Catalase in plants is a heme-coordinated tetrameric protein that primarily disproportionates hydrogen peroxide into water and oxygen. It plays an important role in maintaining cellular concentration of hydrogen peroxide to a level, necessary for all aspects of normal plant growth and development. Except for its recombinant expression in transgenic plants and insect cell line, the protein is yet to be synthesized in its bio-active form in prokaryotic expression system. Attempts made in past for recombinant expression of plant catalase in Escherichia coli consistently resulted in formation of insoluble and inactive aggregates of inclusion body. Here we have shown the specific requirement of a thioredoxin fusion partner, the involvement of trigger factor protein and the low temperature treatment during induction period for synthesis of completely solubilized rice plant catalase-A in recombinant E. coli. Furthermore, the bacteria required the supplementation of ?-aminolevulinic acid to produce bio-active recombinant rice catalase-A. The molecular and biochemical properties of the purified recombinant protein showed the characteristic features of a typical mono-functional plant catalase. These results attest to the usefulness of the present protocol for production of plant catalase using E. coli as heterologous expression system. PMID:21978604
Ray, Mamata; Mishra, Panchanand; Das, Priyanka; Sabat, Surendra Chandra
Investigating catalase activity through hydrogen peroxide decomposition by bacteria biofilms in real time using scanning electrochemical microscopy.
Catalase activity through hydrogen peroxide decomposition in a 1 mM bulk solution above Vibrio fischeri (?-Protebacteria-Vibrionaceae) bacterial biofilms of either symbiotic or free-living strains was studied in real time by scanning electrochemical microscopy (SECM). The catalase activity, in units of micromoles hydrogen peroxide decomposed per minute over a period of 348 s, was found to vary with incubation time of each biofilm in correlation with the corresponding growth curve of bacteria in liquid culture. Average catalase activity for the same incubation times ranging from 1 to 12 h was found to be 0.28 � 0.07 ?mol H2O2/min for the symbiotic biofilms and 0.31 � 0.07 ?mol H2O2/min for the free-living biofilms, suggesting similar catalase activity. Calculations based on Comsol Multiphysics simulations in fitting experimental biofilm data indicated that approximately (3 � 1) � 10(6) molecules of hydrogen peroxide were decomposed by a single bacterium per second, signifying the presence of a highly active catalase. A 2-fold enhancement in catalase activity was found for both free-living and symbiotic biofilms in response to external hydrogen peroxide concentrations as low as 1 nM in the growth media, implying a similar mechanism in responding to oxidative stress. PMID:24328342
Abucayon, Erwin; Ke, Neng; Cornut, Renaud; Patelunas, Anthony; Miller, Douglas; Nishiguchi, Michele K; Zoski, Cynthia G
Catalase activity in macro- and microorganisms as an indicator of biotic stress in coastal waters of the eastern Mediterranean Sea
NASA Astrophysics Data System (ADS)
In this study we examined the activity of catalase in the water column (mainly attributed to planktonic microorganisms) and the activity of catalase and superoxide dismutase (SOD), as well as lipid peroxidation in the midgut gland of the benthic bivalve Donax trunculus as possible indicators of biotic stress. The measurements were performed at stations situated at known contaminated and clean sites in the coastal waters and shores along the Israeli coast (eastern Mediterranean Sea). In the water column, we found that catalase activity was higher in polluted coastal waters than in nearby unpolluted or less-polluted stations. Moreover, there was diurnal periodicity in catalase activity rates which matched the diurnal changes in hydrogen peroxide levels in seawater. Consistent evidence of extracellular catalase activity was found in the seawater sampled. Catalase activity rates in the midgut gland of D. trunculus did not exhibit clear patterns with respect to site (polluted or clean) or season. However, SOD activity and lipid peroxidation measured in the same tissues were good indicators of organic pollution in the coastal waters examined and, among the three stations examined in Haifa Bay, Qiriat Haim was the most polluted.
Angel, D. L.; Fiedler, U.; Eden, N.; Kress, N.; Adelung, D.; Herut, B.
Gram-positive, catalase-positive cocci from dry cured Iberian ham and their enterotoxigenic potential.
Iberian ham is an uncooked, cured meat product ripened under natural uncontrolled conditions for 18 to 24 months. Gram-positive, catalase-positive cocci are the main microbial population in Iberian ham for most of the ripening time. Since some of these organisms are able to produce enterotoxins, adequate characterization and toxicological study are needed. For this, 1,327 gram-positive, catalase-positive cocci, isolated from Iberian hams at different stages and locations, were characterized by physiological and biochemical tests. Selected isolates were further characterized by guanine-cytosine (G+C) content and restriction enzyme analysis of genes coding for 16S rRNA. The toxigenic potential of these organisms was tested with specific DNA gene probes for staphylococcal enterotoxins A, B, C, and D and confirmed by semiquantitative sandwich enzyme immunoassay. The majority of the isolates were identified as Staphylococcus spp. and Micrococcus spp. Non-identified gram-positive, catalase-positive cocci which were moderately halophilic and showed a 42 to 52% G+C content were detected. A great variety of staphylococcal strains were found within the different species at any sampling time. Two strains of Staphylococcus xylosus, one Staphylococcus cohnii strain, and four of the non-identified organisms with 42 to 52% G+C contents hybridized with some of the DNA probes for C and D staphylococcal enterotoxin genes. S. xylosus hybridizing with C-enterotoxin probe reacted with both C and D enterotoxins in the immunological test. In addition, enterotoxin D was confirmed in the nonidentified strains. Some toxigenic organisms were isolated from the final product, posing a health hazard for the consumer.
Rodriguez, M; Nunez, F; Cordoba, J J; Bermudez, E; Asensio, M A
Two temporally synthesized charge subunits interact to form the five isoforms of cottonseed (Gossypium hirsutum) catalase.
Five charge isoforms of tetrameric catalase were isolated from cotyledons of germinated cotton (Gossypium hirsutum L.) seedlings. Denaturing isoelectric focusing of the individual isoforms in polyacrylamide gels indicated that isoforms A (most anodic) and E (most cathodic) consisted of one subunit of different charge, whereas isoforms B, C and D each consisted of a mixture of these two subunits. Thus the five isoforms apparently were formed through combinations of two subunits in different ratios. Labelling cotyledons in vivo with [35S]methionine at three daily intervals in the dark, and translation in vivo of polyadenylated RNA isolated from cotyledons at the same ages, revealed synthesis of two different subunits. One of the subunits was synthesized in cotyledons at all ages studied (days 1-3), whereas the other subunit was detected only at days 2 and 3. This differential expression of two catalase subunits helped explain previous results from this laboratory showing that the two anodic forms (A and B) found in maturing seeds were supplemented with three cathodic forms (C-E) after the seeds germinated. These subunit data also helped clarify our new findings that proteins of isoforms A, B and C (most active isoforms) accumulated in cotyledons of plants kept in the dark for 3 days, then gradually disappeared during the next several days, whereas isoforms D and E (least active isoforms) remained in the cells. This shift in isoform pattern occurred whether seedlings were kept in the dark or exposed to continuous light after day 3, although exposure to light enhanced this process. These sequential molecular events were responsible for the characteristic developmental changes (rise and fall) in total catalase activity. We believe that the isoform changeover is physiologically related to the changeover in glyoxysome to leaf-type-peroxisome metabolism. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. Fig. 5.
Ni, W; Trelease, R N; Eising, R
Catalase activity of cytochrome C oxidase assayed with hydrogen peroxide-sensitive electrode microsensor.
An iron-hexacyanide-covered microelectrode sensor has been used to continuously monitor the kinetics of hydrogen peroxide decomposition catalyzed by oxidized cytochrome oxidase. At cytochrome oxidase concentration ~1 �M, the catalase activity behaves as a first order process with respect to peroxide at concentrations up to ~300-400 �M and is fully blocked by heat inactivation of the enzyme. The catalase (or, rather, pseudocatalase) activity of bovine cytochrome oxidase is characterized by a second order rate constant of ~2�10(2) M(-1)�sec(-1) at pH 7.0 and room temperature, which, when divided by the number of H2O2 molecules disappearing in one catalytic turnover (between 2 and 3), agrees reasonably well with the second order rate constant for H2O2-dependent conversion of the oxidase intermediate F(I)-607 to F(II)-580. Accordingly, the catalase activity of bovine oxidase may be explained by H2O2 procession in the oxygen-reducing center of the enzyme yielding superoxide radicals. Much higher specific rates of H2O2 decomposition are observed with preparations of the bacterial cytochrome c oxidase from Rhodobacter sphaeroides. The observed second order rate constants (up to ~3000 M(-1)�sec(-1)) exceed the rate constant of peroxide binding with the oxygen-reducing center of the oxidized enzyme (~500 M(-1)�sec(-1)) several-fold and therefore cannot be explained by catalytic reaction in the a(3)/Cu(B) site of the enzyme. It is proposed that in the bacterial oxidase, H2O2 can be decomposed by reacting with the adventitious transition metal ions bound by the polyhistidine-tag present in the enzyme, or by virtue of reaction with the tightly-bound Mn2+, which in the bacterial enzyme substitutes for Mg2+ present in the mitochondrial oxidase. PMID:21314602
Bolshakov, I A; Vygodina, T V; Gennis, R; Karyakin, A A; Konstantinov, A A
Temperature dependence of oxygen evolution through catalase-like activity of horseradish peroxidase
NASA Astrophysics Data System (ADS)
By experimental investigations of the temperature dependence of catalase-like activity of horseradish peroxidase in the temperature range 278 328 K, different kinetic profiles for oxygen evolution were found below and above 298 K. Extension of the model is proposed to account for these observations. By numeric simulations of the reaction kinetics at different temperatures, it was found that enhanced evaporation of molecular oxygen from the reaction solution is the main root through which oxygen is lost at elevated temperatures in laboratory conditions.
Popovi?-Bijeli?, A.; Bijeli?, G.; Kolar-Ani?, Lj.; Vukojevi?, V.
Dynamics of the reaction glucose-catalase-glucose oxidase-hydrogen peroxide
NASA Astrophysics Data System (ADS)
Glucose-catalase-glucose oxidase-hydrogen peroxide reaction is one of the few known enzymatic systems studied in vitro in the field of nonlinear chemical dynamics. This reaction belongs to the family of oscillatory enzymatic reactions, which form a natural basis of oscillations in biological systems. A parametric study of dependence on mixing, temperature and initial concentrations of components in a batch stirred reactor was carried out. A newly proposed mathematical model of the reaction conforms to the obtained experimental data. Results of our experiments and simulations hint at further directions of research of non-linear dynamics in this reaction.
?�p, M.; Schreiberov�, L.; Schreiber, I.
X-ray diffraction study of Penicillium Vitale catalase in the complex with aminotriazole
The three-dimensional structure of the enzyme catalase from Penicillium vitale in a complex with the inhibitor aminotriazole was solved and refined by protein X-ray crystallography methods. An analysis of the three-dimensional structure of the complex showed that the inhibition of the enzyme occurs as a result of the covalent binding of aminotriazole to the amino-acid residue His64 in the active site of the enzyme. An investigation of the three-dimensional structure of the complex resulted in the amino-acid residues being more precisely identified. The binding sites of saccharide residues and calcium ions in the protein molecule were found.
Borovik, A. A.; Grebenko, A. I.; Melik-Adamyan, V. R., E-mail: firstname.lastname@example.com [Russian Academy of Sciences, Shubnikov Institute of Crystallography (Russian Federation)
Oxidative stress in Caenorhabditis elegans: protective effects of superoxide dismutase/catalase mimetics.
The lifespan of Caenorhabditis elegans can be extended by the administration of synthetic superoxide dismutase/ catalase mimetics (SCMs) without any effects on development or fertility. Here we demonstrate that the mimetics, Euk-134 and Euk-8, confer resistance to the oxidative stress-inducing agent, paraquat and to thermal stress. The protective effects of the compounds are apparent with treatments either during development or during adulthood and are independent of an insulin/IGF-I-like signalling pathway also known to affect thermal and oxidative stress resistance. Worms exposed to the compounds do not induce a cellular stress response and no detrimental effects are observed. PMID:14677634
Sampayo, James N; Olsen, Anders; Lithgow, Gordon J
Activity of Superoxide Dismutase and Catalase in Fenugreek (Trigonella foenum-graecum) in Response to Carbendazim
Fenugreek (Trigonella foenum-graecum) is an annual herb, used as a spice and traditionally as medicine. Fenugreek finds its uses in treating hyperglycemia, hyperlipidemia and disorders of gastro-intestinal and cardiovascular systems. Fenugreek cultivation in India is affected by fungal diseases like root-rot and damping-off and fungicides like carbendazim are used to overcome these infections. Fungicides play both positive and negative role in plants; fungicides protect plants from diseases and also exert oxidative stress simultaneously. This report is on the response of antioxidants, superoxide dismutase and catalase in fenugreek seeds and plants treated to different concentrations of carbendazim.
Sangeetha, R.
Upregulation of the thioredoxin-dependent redox system during differentiation of 3T3-L1 cells to adipocytes.
Hydrogen peroxide acts as a signaling molecule in early adipogenesis. In differentiating adipocytes, elevated hydrogen peroxide generation is balanced through induction of antioxidant enzymes such as catalase and peroxiredoxins. Thioredoxin reductases (TrxR) and glutathione peroxidases (GPx) are selenoenzymes that constitute part of the major thiol-dependent antioxidant systems in cells. Here we show that the protein levels of cytoplasmic/nuclear TrxR1 and mitochondrial TrxR2 increase in the course of adipocyte differentiation of 3T3-L1 cells together with the TrxR2 substrate thioredoxin 2 (Trx2), resulting in elevated TrxR activity in mature adipocytes. Gene and protein expression of the GPx isoenzyme GPx4 was also stimulated during adipogenesis. Chronic exposure of 3T3-L1 cells to the anti-adipogenic factors tumor necrosis factor ? (TNF-?) or rapamycin during differentiation suppressed TrxR1 and Trx2 upregulation, concomitantly with inhibition of adipogenesis and lipogenesis. In contrast, TNF-? or rapamycin did not affect expression of TrxRs and their Trx substrates in mature adipocytes. These results indicate that upregulation of the thioredoxin-dependent redox system is linked to the development of an adipocyte phenotype. PMID:24516001
Rajalin, Ann-Marie; Micoogullari, Mustafa; Sies, Helmut; Steinbrenner, Holger
Superoxide Dismutase, Catalase, and alpha-Tocopherol Content of Stored Potato Tubers.
Activated oxygen or oxygen free radical mediated damage to plants has been established or implicated in many plant stress situations. The extent of activated oxygen damage to potato (Solanum tuberosum L.) tubers during low temperature storage and long-term storage is not known. Quantitation of oxygen free radical mediated damage in plant tissues is difficult. However, it is comparatively easy to quantitate endogenous antioxidants, which detoxify potentially damaging forms of activated oxygen. Three tuber antioxidants, superoxide dismutase, catalase, and alpha-tocopherol were assayed from four potato cultivars stored at 3 degrees C and 9 degrees C for 40 weeks. Tubers stored at 3 degrees C demonstrated increased superoxide dismutase activities (up to 72%) compared to tubers stored at 9 degrees C. Time dependent increases in the levels of superoxide dismutase, catalase, and alpha-tocopherol occurred during the course of the 40 week storage. The possible relationship between these increases in antioxidants and the rate of activated oxygen production in the tubers is discussed. PMID:16667819
Spychalla, J P; Desborough, S L
Superoxide dismutase, catalase, and. alpha. -tocopherol content of stored potato tubers. [Solanum tuberosum L
Activated oxygen or oxygen free radical mediated damage to plants has been established or implicated in many plant stress situations. The extent of activated oxygen damage to potato (Solanum tuberosum L.) tubers during low temperature storage and long-term storage is not known. Quantitation of oxygen free radical mediated damage in plant tissues is difficult. However, it is comparatively easy to quantitate endogenous antioxidants, which detoxify potentially damaging forms of activated oxygen. Three tuber antioxidants, superoxide dismutase, catalase, and {alpha}-tocopherol were assayed from four potato cultivars stored at 3{degree}C and 9{degree}C for 40 weeks. Tubers stored at 3{degree}C demonstrated increased superoxide dismutase activities (up to 72%) compared to tubers stored at 9{degree}C. Time dependent increases in the levels of superoxide dismutase, catalase, and {alpha}-tocopherol occurred during the course of the 40 week storage. The possible relationship between these increases in antioxidants and the rate of activated oxygen production in the tubers is discussed.
Spychalla, J.P.; Desborough, S.L. (Univ. of Minnesota, St. Paul (USA))
Influence of added catalase on chromosome stability and neoplastic transformation of mouse cells in culture.
The generation of hydrogen peroxide (H2O2) and the derivative free hydroxyl radical (. OH) in cultures of mouse cells grown in the presence of visible light and ambient oxygen was shown previously to be implicated in chromatid damage. Furthermore, chromosome alterations appear to be associated with the spontaneous neoplastic transformation of mouse cells in culture. An attempt was made in this study to reduce the incidence of chromosomal aberrations and delay or prevent the onset of spontaneous neoplastic transformation of freshly isolated mouse cells, both fibroblasts and epidermal keratinocytes, by adding catalase to the culture medium, shielding the cultures from wavelengths less than 500 nm and providing a gas phase of 0-1% O2. These conditions significantly decreased the incidence of chromosomal aberrations in both cell types, and in fibroblasts prevented tumourigenicity in non-irradiated syngeneic mice, and increased latent periods for tumour development in X-irradiated mice. The epidermal keratinocytes were particularly resistant to spontaneous neoplastic transformation under all conditions tested. These observations on the protective effect of extracellular catalase suggest that H2O2, a normal metabolite, and/or the derivative .OH can directly or indirectly produce genetic damage and neoplastic transformation in mouse fibroblasts.
Jones, G. M.; Sanford, K. K.; Parshad, R.; Gantt, R.; Price, F. M.; Tarone, R. E.
Temperature-dependent requirement for catalase in aerobic growth of Listeria monocytogenes F2365.
Listeria monocytogenes is a Gram-positive, psychrotrophic, facultative intracellular food-borne pathogen responsible for severe illness (listeriosis). The bacteria can grow in a wide range of temperatures (1 to 45�C), and low-temperature growth contributes to the food safety hazards associated with contamination of ready-to-eat foods with this pathogen. To assess the impact of oxidative stress responses on the ability of L. monocytogenes to grow at low temperatures and to tolerate repeated freeze-thaw stress (cryotolerance), we generated and characterized a catalase-deficient mutant of L. monocytogenes F2365 harboring a mariner-based transposon insertion in the catalase gene (kat). When grown aerobically on blood-free solid medium, the kat mutant exhibited impaired growth, with the extent of impairment increasing with decreasing temperature, and no growth was detected at 4�C. Aerobic growth in liquid was impaired at 4�C, especially under aeration, but not at higher temperatures (10, 25, or 37�C). Genetic complementation of the mutant with the intact kat restored normal growth, confirming that inactivation of this gene was responsible for the growth impairment. In spite of the expected impact of oxidative stress responses on cryotolerance, cryotolerance of the kat mutant was not affected. PMID:20817809
Azizoglu, Reha Onur; Kathariou, Sophia
Ability of recombinant human catalase to suppress inflammation of the murine lung induced by influenza a.
Influenza A virus pandemics and emerging antiviral resistance highlight the urgent need for novel generic pharmacological strategies that reduce both viral replication and inflammation of the lung. We have previously investigated the therapeutic efficacy of recombinant human catalase (rhCAT) against viral pneumonia in mice, but the protection mechanisms involved were not explored. In the present study, we have performed a more in-depth analysis covering survival, lung inflammation, immune cell responses, production of cytokines, and inflammation signaling pathways in mice. Male imprinting control region mice were infected intranasally with high pathogenicity (H1N1) influenza A virus followed by treatment with recombinant human catalase. The administration of rhCAT resulted in a significant reduction in inflammatory cell infiltration (e.g., macrophages and neutrophils), inflammatory cytokine levels (e.g., IL-2, IL-6, TNF-?, IFN-?), the level of the intercellular adhesion molecule 1 chemokine and the mRNA levels of toll-like receptors TLR-4, TLR-7, and NF-?B, as well as partially maintaining the activity of the antioxidant enzymes system. These findings indicated that rhCAT might play a key protective role in viral pneumonia of mice via suppression of inflammatory immune responses. PMID:24385240
Shi, Xunlong; Shi, Zhihui; Huang, Hai; Zhu, Hongguang; Zhou, Pei; Zhu, Haiyan; Ju, Dianwen
Using liquid crystals for the label-free detection of catalase at aqueous-LC interfaces.
In this study, we developed a simple label-free method for the detection of catalase (CAT) using liquid crystals (LCs). The optical appearance of LCs changed from bright to dark when hydrogen peroxide was in contact with the dodecanal-doped nematic LC, 4-cyano-4'-pentylbiphenyl (5CB). Since hydrogen peroxide can oxidize aldehyde into carboxylic acid, an orientational transition of the LC from the planar to homeotropic state was induced by the self-assembled carboxylate monolayer formed at the aqueous/LC interface. The optical response of LCs exhibited a higher sensitivity to the presence of hydrogen peroxide in an alkaline solution. A new type of LC-based sensor was developed to monitor the presence of CAT in the aqueous phase. Due to the enzymatically catalytic hydrolysis of hydrogen peroxide, the bright-to-dark shift in the optical signal did not take place in the aqueous mixture of hydrogen peroxide and catalase. In contrast, the optical response changed from bright to dark when the mixture in the optical cell was replaced with an aqueous solution of hydrogen peroxide. Considering the optical response of LCs related to the absence and presence of hydrogen peroxide, the aldehyde-doped 5CB might have potential utility in real-time recognition and detection of chemical and biological events associated with hydrogen peroxide. PMID:22138010
Hu, Qiong-Zheng; Jang, Chang-Hyun
Superoxide dismutase mimetics with catalase activity reduce the organ injury in hemorrhagic shock.
Reactive oxygen species (ROS) contribute to the multiple organ failure (MOF) in hemorrhagic shock. Here we investigate the effects of two superoxide dismutase (SOD) mimetics with catalase activity (EUK-8 and EUK-134) on the circulatory failure and the organ injury and dysfunction associated with hemorrhagic shock in the anesthetised rat. Hemorrhage (sufficient to lower mean arterial blood pressure to 45 mmHg for 90 min) and subsequent resuscitation with shed blood resulted (within 4 h after resuscitation) in a delayed fall in blood pressure, liver injury and renal dysfunction as well as pancreatic injury. Treatment of rats on resuscitation with EUK-8 (3 mg/kg i.v. bolus followed by 3 mg/kg/h i.v. infusion) significantly attenuated liver injury, renal dysfunction and pancreatic injury caused by hemorrhage and resuscitation. Administration of EUK-134 (3 mg/kg i.v. bolus followed by 3 mg/kg/h) reduced the liver injury and renal dysfunction (but not the pancreatic injury) caused by hemorrhagic shock. However, neither EUK-8 nor EUK-134 reduced the delayed circulatory failure associated with hemorrhagic shock. Thus, we propose that an enhanced formation of ROS contributes to the MOF in hemorrhagic shock, and that membrane-permeable SOD-mimetics with catalase activity, such as EUK-8 or EUK-134, may represent a novel therapeutic approach for the therapy of hemorrhagic shock. PMID:12353923
Izumi, Maya; McDonald, Michelle C; Sharpe, Martyn A; Chatterjee, Prabal K; Thiemermann, Christoph
Hydrogen peroxide decomposition by a non-heme iron(III) catalase mimic: a DFT study.
Non-heme iron(III) complexes of 14-membered tetraaza macrocycles have previously been found to catalytically decompose hydrogen peroxide to water and molecular oxygen, like the native enzyme catalase. Here the mechanism of this reaction is theoretically investigated by DFT calculations at the (U)B3LYP/6-31G* level, with focus on the reactivity of the possible spin states of the FeIII complexes. The computations suggest that H2O2 decomposition follows a homolytic route with intermediate formation of an iron(IV) oxo radical cation species (L.+FeIV==O) that resembles Compound I of natural iron porphyrin systems. Along the whole catalytic cycle, no significant energetic differences were found for the reaction proceeding on the doublet (S=1/2) or on the quartet (S=3/2) hypersurface, with the single exception of the rate-determining O--O bond cleavage of the first associated hydrogen peroxide molecule, for which reaction via the doublet state is preferred. The sextet (S=5/2) state of the FeIII complexes appears to be unreactive in catalase-like reactions. PMID:17323385
Sicking, Willi; Korth, Hans-Gert; Jansen, Georg; de Groot, Herbert; Sustmann, Reiner
Flow injection catalase activity measurement based on gold nanoparticles/carbon nanotubes modified glassy carbon electrode.
Amperometric flow injection method of hydrogen peroxide analysis was developed based on catalase enzyme (CAT) immobilization on a glassy carbon electrode (GC) modified with electrochemically deposited gold nanoparticles on a multiwalled carbon nanotubes/chitosan film. The resulting biosensor was applied to detect hydrogen peroxide with a linear response range 1.0�10(-7)-2.5�10(-3)M with a correlation coefficient 0.998 and response time less than 10s. The optimum conditions of film deposition such as potential applied, deposition time and pH were tested and the flow injection conditions were optimized to be: flow rate of 3ml/min, sample volume 75?l and saline phosphate buffer of pH 6.89. Catalase enzyme activity was successfully determined in liver homogenate samples of rats, raised under controlled dietary plan, using a flow injection analysis system involving the developed biosensor simultaneously with spectrophotometric detection, which is the common method of enzymatic assay. PMID:22817944
El Nashar, Rasha Mohamed
Catalase and ?-enolase: two novel granulocyte autoantigens in inflammatory bowel disease (IBD)
In IBD, the target antigens of anti-neutrophil cytoplasmic autoantibodies (ANCA) have not been fully identified, which limits the analysis of the diagnostic significance as well as of the possible pathophysiological role of these antibodies. In this study, we identify the target antigens of ANCA in large groups of patients with ulcerative colitis (UC) and Crohn's disease (CD). Apart from antibodies against lactoferrin and bactericidal/permeability-increasing protein (BPI), which have been reported before, antibodies against two novel granulocyte antigens were identified: antibodies against a 57/56-kD doublet were found in 38% of samples from UC patients and in 26% of samples from CD patients, whereas antibodies against a 47-kD protein were found in 10% of samples from UC patients and in 18% of samples from CD patients. Partial purification and amino acid sequence analysis identified the 57-kD protein as catalase and the 47-kD protein as ?-enolase. This study is the first to report catalase and ?-enolase as granulocyte antigens for autoantibodies in IBD.
Roozendaal, C; Zhao, M H; Horst, G; Lockwood, C M; Kleibeuker, J H; Limburg, P C; Nelis, G F; Kallenberg, C G M
Catalase and lipid peroxidation values in serum of Tunisian patients with pemphigus vulgaris and foliaceus.
Pemphigus is an autoimmune disorder resulting from the interaction between autoantibodies and desmoglein. Oxidative stress seems to be responsible for the onset/aggravation of many human diseases. Actually, it is considered as one of the several factors for the etiopathogenesis of pemphigus. The present study aims to evaluate the oxidative state in the sera of pemphigus vulgaris and pemphigus foliaceus patients by assessing lipid peroxidation, proteins oxidation, and antioxidant enzyme activity. This study included 36 pemphigus vulgaris and 42 pemphigus foliaceus patients as well as a group of controls consisting of 78 healthy volunteers. Malondialdehyde levels (p?0.001) and catalase activity (p?0.001) are higher in both groups of patients than in the control group. The two groups of patients showed a nonsignificant decrease in the thiol groups compared with the healthy one. A nonsignificant difference was shown between pemphigus vulgaris and pemphigus foliaceus patients, except for the catalase which shows an increase in the pemphigus vulgaris group. We have also found significant correlations between serum oxidative stress marker levels and serum anti-desmoglein antibody levels in the two pemphigus groups. These findings underline the implication of oxidative stress in the physiopathology of pemphigus by the increase in the autoantibodies' reactivity. PMID:22907559
Abida, Olfa; Ben Mansour, Riadh; Gargouri, Bochra; Ben Ayed, Mourad; Masmoudi, Abderrahmen; Turki, Hamida; Masmoudi, Hatem; Lassoued, Saloua
Sample preparation strategies for quantitative analysis of catalase in red blood cells by elemental mass spectrometry.
A sample preparation strategy for the determination of the Fe-containing enzyme catalase (CAT) by Fe specific monitoring in human erythrocytes has been optimized. For this purpose, the combined use of elemental mass spectrometry (via inductively coupled plasma, ICP-MS), molecular mass spectrometry (via MALDI-TOF) and enzymatic activity measurements has been required. The procedure involved haemoglobin precipitation from cell lysate with a solution of ethanol-chloroform and preconcentration of the supernatant by using a Speed-Vac concentrator. Catalase recoveries of about 88 � 15% could be measured by monitoring the protein enzymatic activity before and after precipitation. Further fractionation of Fe-containing proteins from the preconcentrated extract was achieved by size exclusion chromatography (Superdex 200) with a mobile phase of ammonium acetate (0.05 M, pH 7.4) coupled to ICP-MS (Fe monitoring) and UV/VIS detection (specific absorption of the heme-group at 408 nm). A second dimensional chromatography of the CAT-positive activity fraction was carried out by anion-exchange chromatography (Mono Q 5/50) using for elution a linear gradient of ammonium acetate (0-0.750 M in 15 min). This second step revealed a single Fe-containing species in the chromatogram and permitted the unambiguous characterization of the CAT in such fractions by MALDI-TOF. Column recoveries were evaluated and were quantitative, in terms of Fe bound to protein and CAT activity. PMID:21072355
Mudarra Rubio, A; Montes-Bay�n, M; Blanco-Gonz�lez, E; Sanz-Medel, A
Evidence for separate substrate binding sites for hydrogen peroxide and cumene hydroperoxide (CHP) in the oxidation of ethanol by catalase
The oxidation of ethanol by purified bovine liver catalase (Sigma, C-40) can be supported by H/sub 2/O/sub 2/ or by CHP. The time course of the H/sub 2/O/sub 2/ supported reaction (using glucose/glucose oxidase as the H/sub 2/O/sub 2/ source) was linear for at least one hr, whereas the rate of acetaldehyde formation in the CHP (4.2 mM) supported reaction decreased with time. When catalase was exposed o CHP for 5 min before the addition of ethanol, the rate of CHP supported ethanol oxidation was reduced by more than 90% compared to incubations where the addition of ethanol preceded that of CHP. In the CHP inhibited state, the peroxidative activity of catalase was not restored by further addition of CHP or ethanol; however, addition of fresh catalase yielded its expected activity. Significantly, the CHP inhibited enzyme was equally effective as the untreated enzyme in catalyzing (a) the oxidation of ethanol in the presence H/sub 2/O/sub 2/ supported peroxidative activity as well as catalytic activity by CHP inhibited catalase points to separate binding sites for H/sub 2/O/sub 2/ and CHP in this reaction. Alternatively, CHP may bind adjacent to a common peroxide active site, thereby sterically impeding the binding of CHP - but not of H/sub 2/O/sub 2/ - to this active site.
DeMaster, E.G.; Nagasawa,ss H.T.
Uncaria tomentosa extracts protect human erythrocyte catalase against damage induced by 2,4-D-Na and its metabolites.
The effect of ethanolic and aqueous extracts from leaves and bark of Uncaria tomentosa was studied, with particular attention to catalase activity (CAT - EC. 184.108.40.206). We observed that all tested extracts, at a concentration of 250 ?g/mL were not toxic to erythrocyte catalase because they did not decreased its activity. Additionally, we investigated the protective effect of extracts on changes in CAT activity in the erythrocytes incubated with sodium salt of 2,4-dichlorophenoxyacetic acid (2,4-D-Na) and its metabolites i.e., 2,4-dichlorophenol (2,4-DCP) and catechol. Previous investigations showed that these chemicals decreased activity of erythrocyte catalase (Bukowska et al., 2000; Bukowska and Kowalska, 2004). The erythrocytes were divided into two portions. The first portion was incubated for 1 and 5h at 37�C with 2,4-D-Na, 2,4-DCP and catechol, and second portion was preincubated with extracts for 10 min and then incubated with xenobiotics for 1 and 5h. CAT activity was measured in the first and second portion of the erythrocytes. We found a protective effect of the extracts from U. tomentosa on the activity of catalase incubated with xenobiotics studied. Probably, phenolic compounds contained in U. tomentosa scavenged free radicals, and therefore protected active center (containing -SH groups) of catalase. PMID:22426356
Bukowska, Bo?ena; Bors, Milena; Gulewicz, Krzysztof; Koter-Michalak, Maria
Salicylic acid and salicylic acid sensitive and insensitive catalases in different genotypes of chickpea against Fusarium oxysporum f. sp. ciceri.
Differential expression of catalase isozymes in different genotypes of chickpea resistant genotypes- A1, JG-315, JG-11, WR-315, R1-315, Vijaya, ICCV-15017, GBS-964, GBM-10, and susceptible genotypes- JG-62, MNK, ICCV-08321, ICCV-08311, KW-104, ICCV-08123, ICC-4951, ICC-11322, ICC-08116 for wilt disease caused by Fusarium oxysporum. f. sp. ciceri (Foc) was analyzed. Salicylic acid (SA) and H2O2 concentrations were determined in control as well as in plants infected with F. ciceri and found that the high and low levels of salicylic acid and H2O2 in resistant and susceptible genotypes of chickpea respectively. Catalase isozyme activities were detected in the gel and found that no induction of new catalases was observed in all the resistant genotypes and their some of the native catalase isozymes were inhibited; whereas, induction of multiple catalase isozymes was observed in all the screened susceptible genotypes and their activities were not inhibited upon Foc or SA treatments. The above results support the possible role of these isozymes as a marker to identify which genotype of chickpea is expressing systemic acquired resistance. PMID:24431522
Gayatridevi, S; Jayalakshmi, S K; Mulimani, V H; Sreeramulu, K
Scavenging of free radicals in gas-phase mainstream cigarette smoke by immobilized catalase at filter level.
Catalase is well known as capable of inducing the decomposition of H(2)O(2). In this study, a kind of immobilized catalase (entrapped in cross-linked chitosan beads) was dispersed in conventional acetate filter as an antioxidant additive. Quantitative estimation of the free radicals in mainstream cigarette smoke (MCS) was performed to address the effect of this modified filter. It was found that the levels of PBN adduct and NO(*)/NO(2)(*) associated with the gas-phase mainstream cigarette smoke (GPCS) were efficiently decreased by approximately 40% through catalase filtering. Besides, the modified filter was found to lower the MCS-induced adverse biological effects including lipid peroxidation and mutagenicity. This was proved to be substantially attributed to the catalase-dependent breakdown of NO(*), which was stimulated by some of peroxides (most probably being H(2)O(2)), the dismutation products of tar particulate matters (TPM). These results highlighted a promising approach to reduce the smoking-associated health risks to passive smokers. Moreover, the mechanisms of catalase filtering may be helpful for the development of appropriate immobilized enzyme systems to be applied for reducing health risks associated with gaseous pollutants. PMID:18344119
Lu, Xin; Hua, Zhaozhe; Du, Guocheng; Ma, Xiaolong; Cao, Jianhua; Yang, Zhanping; Chen, Jian
Choline Promotes Nicotinic Receptor ?4 + ?2 Up-regulation*
Neuronal nicotinic acetylcholine receptors (nAChR) composed of ?4 + ?2 subunits, the high affinity nicotine-binding site in the mammalian brain, up-regulate in response to chronic nicotine exposure. The identities of endogenous mediators of this process are unknown. We find that choline also up-regulates ?4 + ?2 nAChRs stably expressed by HEK293 cells as measured by increased [3H]epibatidine density. Choline-mediated up-regulation is dose-dependent and corresponds with an increase in ?2 subunit protein expression. The choline kinase inhibitor hemicholinium-3 inhibits ?60% of choline-mediated up-regulation revealing both an HC3-dependent and -independent pathway. Furthermore, choline-mediated up-regulation is not additive with up-regulation agents such as nicotine, but it is additive with weaker promoters of the up-regulation process. When co-applied with the pro-inflammatory cytokine tumor necrosis factor ?, choline-mediated up-regulation is increased further through a mechanism that includes an increase in both ?4 and ?2 protein expression, and this is inhibited by the p38 MAPK inhibitor SB202190. These findings extend the view that up-regulation of ?4 + ?2 nAChRs is a normal physiological response to altered metabolic and inflammatory conditions.
Gahring, Lorise C.; Vasquez-Opazo, Gustavo A.; Rogers, Scott W.
Evolutionary relationship of plant catalase genes inferred from exon-intron structures: isozyme divergence after the separation of monocots and dicots
In order to understand the molecular evolution of catalase genes in higher plants, we compared the exon-intron structures\\u000a of 12 genomic sequences from six plant species. It was assumed that the putative single primordial catalase gene had seven\\u000a introns, because only those catalase genes having this structure are found in the monocotyledonae and dicotyledonae classes.\\u000a After the evolutionary divergence of
M. Iwamoto; M. Maekawa; A. Saito; H. Higo; K. Higo
Isolation of Catalase-NegativeListeria monocytogenesStrains from Listeriosis Patients and Their Rapid Identification by Anti-p60 Antibodies and\\/or PCR
Two catalase-negative Listeria monocytogenes serovar 1\\/2b strains were isolated from listeriosis patients in 1995 in Germany. The infections appeared in individuals from different cities at different seasons and were caused byL. monocytogenesstrains of different clonal types. In particular, the catalase reaction of one strain isolatedfrombloodwasconsistentlynegative,whereasthisreactionwasonlyreversiblyblockedwhenthestrain was freshly isolated from asciticfluid. After subculturing, the catalase-positive reaction was restored. Initially, identification of
ANDREAS BUBERT; JOACHIM RIEBE; NORBERT SCHNITZLER; ARNO SCHONBERG; WERNER GOEBEL; ANDPETER SCHUBERT; E. Merck
Catalase and alternative oxidase cooperatively regulate programmed cell death induced by beta-glucan elicitor in potato suspension cultures.
In potato (Solanum tuberosum L.) suspension cells, the expression of the gene encoding alternative oxidase (AOX) and H2O2 accumulation were induced by treatment with beta-glucan elicitor. The inhibition of catalase activity enhanced both AOX mRNA expression and the production of H2O2, whereas the ascorbate peroxidase inhibitor did not have any effect on these responses. Simultaneous inhibition of catalase and AOX activities in elicited cells dramatically increased H2O2 accumulation, leading to the disruption of mitochondrial membrane potential (deltapsi(m)) and programmed cell death (PCD). The results demonstrate, for the first time, that not only AOX but also catalase plays a central role in the suppression of mitochondrial deltapsi(m) breakdown and PCD induced by beta-glucan elicitor. PMID:15480753
Mizuno, Masashi; Tada, Yasuomi; Uchii, Kimitaka; Kawakami, Sachiko; Mayama, Shigeyuki
Characterization of the katG gene encoding a catalase-peroxidase required for the isoniazid susceptibility of Mycobacterium tuberculosis.
The isoniazid susceptibility of Mycobacterium tuberculosis is mediated by the product of the katG gene which encodes the heme-containing enzyme catalase-peroxidase. In this study, the chromosomal location of katG has been established and its nucleotide sequence has been determined so that the primary structure of catalase-peroxidase could be predicted. The M. tuberculosis enzyme is an 80,000-dalton protein containing several motifs characteristic of peroxidases and shows strong similarity to other bacterial catalase-peroxidases. Expression of the katG gene in M. tuberculosis, M. smegmatis, and Escherichia coli was demonstrated by Western blotting (immunoblotting). Homologous genes were detected in other mycobacteria, even those which are naturally insensitive to isoniazid. Images
Heym, B; Zhang, Y; Poulet, S; Young, D; Cole, S T
Effects of superoxide dismutase/catalase mimetics on life span and oxidative stress resistance in the housefly, Musca domestica.
The purpose of this study was to determine whether superoxide dismutase/catalase mimetics lengthen the life span of the housefly, Musca domestica, as previously demonstrated for the nematode Caenorhabditis elegans. Various concentrations of Eukarion-8 or Eukarion-134 were administered via the drinking water and the effects on the life span of the flies and amounts of protein carbonyls were determined under normoxic and hyperoxic conditions. These SOD/catalase mimetics neither extended the life span of the flies nor attenuated the protein carbonyl content under normoxic conditions and shortened life span under hyperoxic conditions. Thus, the effect of these SOD/catalase mimetics on the life span of animals seem to be species-specific. PMID:12031907
Bayne, Anne-C�cile V; Sohal, Rajindar S
Effects of molecular oxygen on detection of superoxide radical with nitroblue tetrazolium and on activity stains for catalase
The usual method of staining polyacrylamide gel electropherograms for superoxide dismutase activity utilizes a photochemical flux of O/sub 2//sup -/ to reduce nitroblue tetrazolium. Superoxide dismutases intercept O/sub 2//sup -/, preventing formazan production and thus causing achromatic bands. In the presence of H/sub 2/O/sub 2/, catalase also yield achromatic bands during this staining procedure. This is due to local elevation of pO/sub 2/ by the catalytic decomposition of H/sub 2/O/sub 2/. O/sub 2/, in turn, inhibits the reduction of the tetrazolium by O/sub 2//sup -/. This phenomenon provides a new activity stain for catalase. A previously described activity stain for catalase has also been reexamined and significantly improved.
Clare, D.A.; Duong, M.N.; Darr, D.; Archibald, F.; Fridovich, I.
A kinetic method for para -nitrophenol determination based on its inhibitory effect on the catalatic reaction of catalase
The inhibitory effect of para-nitrophenol on the catalytic reaction of catalase was investigated. Michaelis-Menten kinetic parameters were determined from\\u000a Lineweaver-Burk plots obtained in the absence or in the presence of the inhibitor. The inhibitor pattern, revealed by the\\u000a Lineweaver-Burk plots, suggested a fully mixed inhibition mechanism. Spectrophotometric monitoring of the indicator reaction:\\u000a $$H_2 O_2 \\\\xrightarrow{{catalase,para - nitrophenol}}H_2 O + \\\\tfrac{1}{2}O_2
Claudia Mure?anu; Lucian Copolovici; Florina Pog�acean
Erythropoietin upregulation in pulmonary arterial hypertension
Abstract The pathophysiologic alterations of patients with pulmonary arterial hypertension (PAH) are diverse. We aimed to determine novel pathogenic pathways from circulating proteins in patients with PAH. Multianalyte profiling (MAP) was used to measure 90 specifically selected antigens in the plasma of 113 PAH patients and 51 control patients. Erythropoietin (EPO) functional activity was assessed via in vitro pulmonary artery endothelial cell networking and smooth muscle cell proliferation assays. Fifty-eight patients had idiopathic PAH, whereas 55 had other forms of PAH; 5 had heritable PAH, 18 had connective tissue disease (15 with scleroderma and 3 with lupus erythematosis), 13 had portopulmonary hypertension, 6 had PAH associated with drugs or toxins, and 5 had congenital heart disease. The plasma-antigen profile of PAH revealed increased levels of several novel biomarkers, including EPO. Immune quantitative and histochemical studies revealed that EPO not only was significantly elevated in the plasma of PAH patients but also promoted pulmonary artery endothelial cell network formation and smooth muscle cell proliferation. MAP is a hypothesis-generating approach to identifying novel pathophysiologic pathways in PAH. EPO is upregulated in the circulation and lungs of patients with PAH and may affect endothelial and smooth muscle cell proliferation.
Catalase-positive microperoxisomes in rat soleus and extensor digitorum longus muscle fiber types
NASA Technical Reports Server (NTRS)
The size, distribution, and content of catalase-reactive microperoxisomes were investigated cytochemically in three types of muscle fibers from the soleus and the extensor digitorum longus (EDL) of male rats. Muscle fibers were classified on the basis of the mitochondrial content and distribution, the Z-band widths, and the size and shape of myofibrils as the slow-twitch oxidative (SO), the fast-twitch oxidative glycolytic (FOG), and the fast-twitch glycolytic (FG) fibers. It was found that both the EDL and soleus SO fibers possessed the largest microperoxisomes. A comparison of microperoxisome number per muscle fiber area or the microperoxisome area per fiber area revealed following ranking, starting from the largest number and the area-ratio values: soleus SO, EDL SO, EDL FOG, and EDL FG.
Riley, Danny A.; Bain, James L. W.; Ellis, Stanley
Spectroscopic Investigations on the Interaction between Carbon Nanotubes and Catalase on Molecular Level.
The interactions between well-dispersed multiwalled carbon nanotubes (MWCNTs) and catalase (CAT) were investigated. The activity of CAT was inhibited with the addition of MWCNTs. After deducting the inner filter effect, the fluorescence spectra revealed that the tryptophan (Trp) residues were exposed and the fluorescence intensities of CAT increased with the increase in the MWCNTs concentration. At the same time, the environment of the Trp residues became more hydrophobic. The results of UV-vis absorption spectroscopy and CD spectra indicated that the secondary structure of CAT had been changed, and the amino acid residues were located in a more hydrophobic environment. Meanwhile, the UV-vis spectra indicated that the conformation of the heme porphyrin rings was changed. The microenvironment of CAT activity sites may be interfered by MWCNTs. This research showed that MWCNTs could not only contribute to the conformational changes of protein but also change the enzyme function. PMID:24616245
Guan, Jin; Dai, Jingping; Zhao, Xingchen; Liu, Chunhua; Gao, Canzhu; Liu, Rutao
Cu(II)-disulfide complexes display simultaneous superoxide dismutase- and catalase-like activities.
Superoxide is a potentially toxic by-product of cellular metabolism. We have addressed here the in vitro ability of complexes formed between copper(II) ions and various biologically-occurring disulfides (RSSR: oxidized glutathione, cystine, homocystine and ?-lipoic acid) to react with superoxide. The studied complexes were found to react with superoxide (generated by a xanthine/xanthine oxidase system) at rate constants (kCu(II)-RSSR) close to 10(6)M(-1)s(-1), which are three orders of magnitude lower than that reported for superoxide dismutase (SOD) but comparable to that of several other copper-containing complexes reported as SOD mimetics. The interaction between the tested Cu(II)-RSSR and superoxide, led to the generation and recovery of concentrations of hydrogen peroxide and oxygen that were, respectively, below and above those theoretically-expected from a sole SOD mimetic action. Interestingly, oxygen was generated when the Cu(II)-RSSR complexes were directly incubated with hydrogen peroxide. Taken together, these results reveal that the Cu(II)-RSSR complexes not only have the capacity to dismutate superoxide but also to simultaneously act like catalase mimetic molecules. When added to superoxide-overproducing mitochondria (condition attained by its exposure to diclofenac), three of the tested complexes were able (2-4?M), not only to totally restore, but also to lower below the basal level the mitochondrial production of superoxide. The present study is first in reporting on the potential of Cu(II)-disulfide complexes to act as SOD and catalase like molecules, suggesting a potential for these types of molecules to act as such under physiological and/or oxidative-stress conditions. PMID:24103366
Aliaga, Margarita E; Andrade-Acu�a, Daniela; L�pez-Alarc�n, Camilo; Sandoval-Acu�a, Cristi�n; Speisky, Hern�n
The effect of intracellular delivery of catalase and antisense oligonucleotides to NF-kappaB using albumin microcapsules in the endotoxic shock model.
Microencapsulated (MC) catalase has been shown to inhibit H(2)O(2) and tumor necrosis factor (TNF) in vitro after endotoxin stimulation. It is the purpose of this study to determine whether MC catalase improves pro-inflammatory cytokine inhibition and mortality in an endotoxic shock model in vivo. We also examined whether MC catalase and antisense oligonucleotides (ASO) to nuclear factor kappaB (NF-kappaB) together improved survival by inhibiting pro-inflammatory cytokines using different mechanisms. Methods: Albumin microcapsules containing catalase and ASO to NF-kappaB were prepared 2-7 microm in size by using a B�chi spray dryer. Progressively increasing doses of MC catalase, MC ASO to NF-kappaB, and the combination were given to rats before the administration of Escherichia coli endotoxin. Results demonstrated 60% survival in rats given 15 mg/kg MC catalase, 70% survival with 20 mg/kg MC ASO NF-kappaB, and 80% survival with the combination. TNF was inhibited by 53% in the MC catalase group 4 h after endotoxin administration, 43% in the ASO NF-kappaB group, and 78% in the combination group compared to controls. In conclusion, this study demonstrates the effectiveness of MC intracellular delivery of the naturally occurring antioxidant catalase in improving animal survival. The addition of ASO to NF-kappaB improved both cytokine inhibition and animal survival in endotoxic shock. PMID:19845486
Siwale, Rodney C; Oettinger, Carl W; Addo, Richard; Siddig, Aladin; D'Souza, Martin J
Characterization of a pathogen-induced potato catalase and its systemic expression upon nematode and bacterial infection.
We have isolated a cDNA encoding a catalase (Cat2St) by differential screening of a cDNA library constructed from potato roots infected with the cyst nematode Globodera pallida. Expression analysis confirmed the local induction of Cat2St and showed that it was highest at the adult stage of the parasite. It also revealed that Cat2St was induced in uninfected roots, stems, and leaves of infected plants. Localized and systemic induction of Cat2St was also observed upon root-knot nematode (Meloidogyne incognita) and root bacteria (Erwinia carotovora, Corynebacterium sepedonicum) infections. Based on sequence and expression analysis, Cat2St was found to belong to the recently described class II of dicotyledonous catalases, suggesting that these catalase isoforms could also be pathogen induced. Plant-parasitic nematodes are known to induce, in the roots of their hosts, highly metabolic feeding cells that function as nutritional sinks. Whereas the local induction of Cat2St is probably a consequence of an oxidative stress of metabolic nature, the systemic induction of Cat2St shows striking similarities with the induction of systemic acquired resistance (SAR) genes. The possible role of catalase in compatible plant-pathogen interactions is discussed. PMID:7655060
Niebel, A; Heungens, K; Barthels, N; Inz�, D; Van Montagu, M; Gheysen, G
Catalase-positive cocci in fermented sausage: Variability due to different pork breeds, breeding systems and sausage production technology
The aim of this study was to compare the ecology of catalase-positive cocci (CPC) present in traditional fermented sausages produced using different breeds of pork, each of which was raised in two different environments and processed using two different technologies. Semi-quantitative molecular methods were used to determine bacterial identities. Almost all fermentations were characterised by a significant increase in CPC
Lucilla Iacumin; Marisa Manzano; Giuseppe Comi
Preliminary Study on Glucose Oxidase�Catalase Enzyme System to Control the Browning of Apple and Pear Pur�es
Oxygen is a key factor in fruit pur�es browning. The use of a commercial glucose oxidase�catalase enzyme system (GOX) for oxygen removal and browning control of Golden Delicious apple and Kaiser pear pur�es during storage has been investigated. The effect of ascorbic acid and peroxidase has also been tested and results compared with control (no treatment). Colour change was quantified
G. P. Parpinello; F. Chinnici; A. Versari; C. Riponi
Cationization of catalase, peroxidase, and superoxide dismutase. Effect of improved intraarticular retention on experimental arthritis in mice.
Several enzymes and other proteins were made cationic either by coupling to polylysine or by shielding of anionic sites. These cationic proteins, all having an isoelectric point greater than 8.5 exhibited excellent retention in articular structures when injected in mouse knee joints. Autoradiography and histochemistry showed that cationic forms of catalase, superoxide dismutase, and horseradish peroxidase were firmly retained by synovial and cartilaginous tissues. The half-life of these enzymes in the joint is thus significantly extended compared with native enzymes. The native enzymes and their cationic derivatives were tested for antiinflammatory properties in mice, using antigen-induced arthritis and zymosan-induced arthritis. It was found that injection of cationic catalase or peroxidase induced a marked suppression of some parameters of the inflammatory response in both types of arthritis, as measured by 99m technetium pertechnetate uptake and leakage of 125I-labeled albumin. Native catalase and peroxidase were less, or not at all effective. Cationic superoxide dismutase or cationic nonenzyme proteins did not suppress inflammation. The observed suppression of two different types of inflammation (an immune and a nonimmune arthritis) by catalase and peroxidase suggests that elimination of peroxides contributes to the suppression of an inflammatory response. We would hypothesize that cationic enzymes offer the possibility for investigating the mechanisms of inflammation and, in addition, might be interesting from a therapeutical point of view. Images
Schalkwijk, J; van den Berg, W B; van de Putte, L B; Joosten, L A; van den Bersselaar, L
Low erythrocyte catalase enzyme activity is correlated with high serum total homocysteine levels in tunisian patients with acute myocardial infarction
Background An imbalance between pro-oxidants and antioxidant systems has been suggested to be implicated in the physiopathology of acute myocardial infarction (AMI). We aimed to evaluate the antioxidant capacity in Tunisian patients and to assess the possible relationship between erythrocyte catalase enzyme activity and hyperhomocysteinaemia. Methods 108 patients with AMI and 81 healthy subjects were enrolled in this study. Catalase erythrocyte enzyme activity was determined spectrophotometrically whereas �total antioxidant status� (TAS) concentration was measured by a commercially available method. Serum total homocysteine (tHcy) level was determined by a fluorescence polarization immunoassay (FPIA). Lipid peroxidation was measured with a fluorimetric method as �thiobarbituric acid reactive substances� (TBARS). Results Compared with healthy subjects, patients with AMI had significantly lower catalase activity (P<0.001), TAS concentrations (P<0.001), and significantly higher serum tHcy (P<0.001) and TBARS levels (P<0.001). Erythrocyte catalase enzyme activity was negatively correlated with serum tHcy and TBARS while serum tHcy and TBARS were in positive correlation. Furthermore, the unbalance between pro-oxidants and antioxidants seems to be more aggravated in patients with Q wave AMI compared to patients with non-Q wave AMI. Conclusion Our results suggest the involvement of hyperhomocysteinaemia in the drop of erythrocyte catalase activity related to myocardial ischemia reperfusion. Hyperhomocysteinaemia may increase the myocardial wall dysfunction under ischemia reperfusion by excessive production of reactive oxygen species which is made evident by increased lipid peroxidation. Virtual slides The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1623509866881834
Mitochondrial mutations contribute to HIF1? accumulation via increased reactive oxygen species and upregulated PDK2 in head and neck squamous cell carcinoma
Purpose Mitochondrial mutations have been identified in head and neck squamous cell carcinoma (HNSCC), but the pathways by which phenotypic effects of these mutations are exerted remain unclear. Previously, we found that mitochondrial ND2 mutations in primary HNSCC increased reactive oxygen species (ROS) and conferred an aerobic, glycolytic phenotype with HIF1? accumulation and increased cell growth. The purpose of present study was to examine the pathways relating these alterations. Experimental Design Mitochondrial mutant and wild-type ND2 constructs were transfected into oral keratinocyte immortal cell line OKF6 and head and neck cancer cell line JHU-O19 and established transfectants. The protein levels of HIF1?, pyruvate dehydrogenease (PDH), phospho-PDH, and pyruvate dehydrogenease kinase (PDK) 2, together with ROS generation, were compared between the mutant and wild type. Meanwhile, the effects of small molecule inhibitors targeting PDK2, and mitochondrial targeted catalase, were evaluated on the ND2 mutant transfectants. Results We determined that ND2 mutant downregulated PDH expression via upregulated PDK2, with an increase in phospho-PDH. Inhibition of PDK2 with dichloroacetate decreased HIF1? accumulation and reduced cell growth. Extracellular treatment with hydrogen peroxide, a ROS mimic, increased PDK2 expression and HIF1? expression, and introduction of mitochondrial targeted catalase decreased mitochondrial mutation mediated PDK2 and HIF1? expression and suppressed cell growth. Conclusions Our findings suggest that mitochondrial ND2 mutation contributes to HIF1? accumulation via increased ROS production, upregulation of PDK2, attenuating PDH activity, thereby increasing pyruvate, resulting in HIF1? stabilization. This may provide insight into a potential mechanism by which mitochondrial mutations contribute to HNSCC development.
Sun, Wenyue; Zhou, Shaoyu; Chang, Steven S.; McFate, Thomas; Verma, Ajay; Califano, Joseph A.
Oxidative stress upregulates PDCD4 expression in patients with gastric cancer via miR-21.
Reactive oxygen species (ROS) plays a key role in carcinogenesis by aberrantly inducing signaling networks that initiatiate tumorigenesis and stimulate tumor progression. MicroRNAs (miRNAs) comprise a novel class of endogenous, small, noncoding RNAs that negatively regulate approximately 30% of the genes in a cell via degradation or translational inhibition of their target mRNAs. However, the effects of ROS on miRNAs expression and the role of miRNAs in ROS-mediated injury on carcinogenesis are uncertain. Using UV spectrophotometry and enzyme-linked immunosorbent assay (ELISA), we examined tissues from human gastric cancers and tissues adjascent to gastric cancer and normal gastric tissues and found that total anti-oxidation competence (T-AOC), superoxide dismutase (SOD) and catalase (CAT) concentrations were lower in gastric cancer patients compared to the control subjects, while the concentrations of DNA oxidative damage product 8-oxo-deoxyguanosine (8-OHdG) was higher. To determine the potential role of miRNA in gastric carcinogenesis, real-time quantitative polymerase chain reaction (QPCR) analysis was performed. We found that human 8-oxoguanine DNA N-glycosylase 1 (hOGG1) mRNA and miR-21 expression were significantly upregulated in gastric cancer tissues than in the adjacent normal gastric tissues. Furthermore, the expression of programmed cell death 4 protein (PDCD4) in gastric cancer tissues was significantly lower than in adjacent normal gastric tissues. The expression of miR-21 and PDCD4 was highly correlated with the degree of differentiation, tumor staging, local lymphatic node metastasis and remote metastasis. Expression of miR-21 was negatively correlated with T-AOC, SOD and CAT, but positively correlated with 8-OHdG and hOGG1mRNA. In addition, the relative expression of PDCD4 was negatively correlated with miR-21. These results suggest that the defensive balance of oxidation and antioxidant system in patients with GC was impaired, resulting in enhanced oxidative tissue injury, which may directly contribute to gastric carcinogenesis. Thus we conclude that ROS promotes gastric carcinogenesis via upregulating miR-21 expression which in turn down-regulates the expression of PDCD4 in gastric cancer cells. PMID:23888942
Tu, Honglei; Sun, Haibing; Lin, Yan; Ding, Jie; Nan, Kejun; Li, Zongfang; Shen, Qiang; Wei, Yongchang
Failure of catalase to protect against aflatoxin B{sub 1}-induced mouse lung tumorigenicity
The carcinogenic mycotoxin aflatoxin B{sub 1} (AFB{sub 1}) induces 8-hydroxy-2'-deoxyguanosine (8-OHdG) formation in mouse lung, an effect that can be prevented by treatment with polyethylene glycol-conjugated catalase (PEG-CAT). G {yields} T transversion mutation in K-ras, an early event in AFB{sub 1}-induced mouse lung carcinogenesis, is thought to result from AFB{sub 1}-8,9-exo-epoxide binding to DNA to form AFB{sub 1}-N{sup 7}-guanine, but may also result from formation of 8-OHdG. Therefore, oxidative DNA damage may be important in AFB{sub 1} carcinogenicity. The objective of this study was to determine whether PEG-CAT would prevent AFB{sub 1} tumorigenicity. Mouse lung tumorigenesis was assessed following treatment of female A/J mice with 300 kU/kg PEG-CAT ip and/or 50 mg/kg AFB{sub 1}. Mice were killed 7 months post-treatment and tumors greater than 1 mm in diameter were excised. Unexpectedly, the mean number of tumors per mouse in the PEG-CAT + AFB{sub 1} group (8.81 {+-} 3.64, n = 47) was greater than that of the group treated with AFB{sub 1} alone (7.05 {+-} 3.45, n = 42) (P < 0.05). The tumors obtained from mice treated with PEG-CAT + AFB{sub 1} were larger than those from mice treated with AFB{sub 1} alone (P < 0.05). There was no difference in K-ras exon 1 mutation spectrum or in the histological diagnosis of tumors between AFB{sub 1} and PEG-CAT + AFB{sub 1} groups (P > 0.05). In vitro incubation with mouse liver catalase (CAT) resulted in conversion of [{sup 3}H]AFB{sub 1} into a DNA-binding species, a possible explanation for the results observed in vivo. These results demonstrate that PEG-CAT is not protective against AFB{sub 1} carcinogenicity in mouse lung despite preventing DNA oxidation.
Guindon, Katherine A. [Department of Pharmacology and Toxicology, Queen's University, Kingston, Ontario, K7L 3N6 (Canada); Foley, Julie F.; Maronpot, Robert R. [National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709 (United States); Massey, Thomas E. [Department of Pharmacology and Toxicology, Queen's University, Kingston, Ontario, K7L 3N6 (Canada)], E-mail: firstname.lastname@example.com
HDAC inhibition upregulates the expression of angiostatic ADAMTS1
HDAC inhibitors are promising anticancer agents that induce cell cycle arrest and apoptosis. However, the role of HDACs in cancer progression, such as angiogenesis and metastasis, remains largely unexplored. Among various HDAC inhibitors, we demonstrate that TSA and SAHA upregulated the expression of angiostatic ADAMTS1 in A549 cells. HDAC6 inhibitor tubacin, and knockdown of HDAC6, also lead to ADAMTS1 upregulation.
Chia-Wei Chou; Ching-Chow Chen
Expression of a cloned sweet potato catalase SPCAT1 alleviates ethephon-mediated leaf senescence and H?O? elevation.
In this report a full-length cDNA, SPCAT1, was isolated from ethephon-treated mature L3 leaves of sweet potato. SPCAT1 contained 1479 nucleotides (492 amino acids) in its open reading frame, and exhibited high amino acid sequence identities (ca. 71.2-80.9%) with several plant catalases, including Arabidopsis, eggplant, grey mangrove, pea, potato, tobacco and tomato. Gene structural analysis showed that SPCAT1 encoded a catalase and contained a putative conserved internal peroxisomal targeting signal PTS1 motif and calmodulin binding domain around its C-terminus. RT-PCR showed that SPCAT1 gene expression was enhanced significantly in mature L3 and early senescent L4 leaves and was much reduced in immature L1, L2 and completely yellowing senescent L5 leaves. In dark- and ethephon-treated L3 leaves, SPCAT1 expression was significantly enhanced temporarily from 0 to 24h, then decreased gradually until 72h after treatment. SPCAT1 gene expression levels also exhibited approximately inverse correlation with the qualitative and quantitative H(2)O(2) amounts. Effector treatment showed that ethephon-enhanced SPCAT1 expression was repressed by antioxidant reduced glutathione, NADPH oxidase inhibitor diphenylene iodonium (DPI), calcium ion chelator EGTA and de novo protein synthesis inhibitor cycloheximide. These data suggest that elevated reactive oxygen species H(2)O(2), NADPH oxidase, external calcium influx and de novo synthesized proteins are required and associated with ethephon-mediated enhancement of sweet potato catalase SPCAT1 expression. Exogenous application of expressed catalase SPCAT1 fusion protein delayed or alleviated ethephon-mediated leaf senescence and H(2)O(2) elevation. Based on these data we conclude that sweet potato SPCAT1 is an ethephon-inducible peroxisomal catalase, and its expression is regulated by reduced glutathione, DPI, EGTA and cycloheximide. Sweet potato catalase SPCAT1 may play a physiological role or function in cope with H(2)O(2) homeostasis in leaves caused by developmental cues and environmental stimuli. PMID:21893366
Chen, Hsien-Jung; Wu, Sin-Dai; Huang, Guan-Jhong; Shen, Che-Yu; Afiyanti, Mufidah; Li, Wei-Jhen; Lin, Yaw-Huei
Multifrequency EPR Studies of Manganese Catalases Provide a Complete Description of Proteinaceous Nitrogen Coordination
Pulse electron paramagnetic resonance (EPR) spectroscopy is employed at two very different excitation frequencies, 9.77 and 30.67 GHz, in the study of the nitrogen coordination environment of the Mn(III)Mn(IV) state of the dimanganese-containing catalases from Lactobacillus plantarum and Thermus thermophilus. Consistent with previous studies, the lower-frequency results reveal one unique histidine nitrogen-Mn cluster interaction. For the first time, a second, more strongly hyperfine-coupled 14N atom is unambiguously observed through the use of higher frequency/higher field EPR spectroscopy. The low excitation frequency spectral features are rationalized as arising from the interaction of a histidine nitrogen that is bound to the Mn(IV) ion, while the higher excitation frequency features are attributed to the histidine nitrogen bound to the Mn(III) ion. These results allow for the computation of intrinsic hyperfine coupling constants, which range from 2.2 to 2.9 MHz, for sp2-hybridized nitrogens coordinating equatorially to high-valent Mn ions. The relevance of these findings is discussed in the context of recent results from analogous higher frequency EPR studies of the Mn cluster in photosystem II and other exchange-coupled transition metal-containing systems.
Stich, Troy A.; Whittaker, James W.; Britt, R. David
Antioxidant effects of black rice extract through the induction of superoxide dismutase and catalase activities.
Our ex vivo study revealed that BRE had significantly stronger ability to inhibit LDL oxidation than white rice extract (WRE). The purpose of this study was to investigate whether black rice extract (BRE) supplementation might ameliorate oxidative stress and enhance antioxidant enzyme activities in HepG2 cells and in C57BL/6 mice. In the cellular study, superoxide anions (O2*-) and reactive oxygen species (ROS) in the BRE group were significantly suppressed. The BRE group also showed significant increases in superoxide dismutase (SOD) and catalase (CAT) activities by 161.6% and 73.4%, respectively. The major components responsible for the free-radical-scavenging and antioxidative properties might be cyanidin-3-O-glucoside chloride and peonidin-3-O-glucuside chloride. In the animal study, male C57BL/6 mice were divided into three groups (control, BRE, and WRE). Plasma HDL-cholesterol was significantly higher, and thiobarbituric, acid-reactive substances were significantly lower in the BRE group, whereas plasma levels of total cholesterol and triglyceride were not affected by BRE supplementation. Increased hepatic SOD and CAT activities were observed in BRE-treated mice as compared to the control mice. However, no changes were detected for the protein expression of antioxidant enzymes by Western blot analysis. Our data suggest that antioxidative effects exerted by BRE are mediated through decreases in free-radical generation as well as increases in SOD and CAT activities both in vitro and in vivo. PMID:17120934
Chiang, An-Na; Wu, Hua-Lin; Yeh, Hung-I; Chu, Chi-Shuen; Lin, Hui-Chiao; Lee, Wei-Chin
Biopharmacology of Enzyme Conjugates: Vasoprotective Activity of Supramolecular Superoxide Dismutase-Chondroitin Sulfate-Catalase Derivative
Bienzyme conjugate was obtained by the covalent connection of superoxide dismutase with catalase through endothelial glycocalyx glycosaminoglycan � chondroitin sulfate (SOD-CHS-CAT). This SOD-CHS-CAT conjugate has vasoprotective activity in respect to platelet interactions, tonus of the ring arterial fragment of a rat blood vessel, as well as normalization of hemodynamic parameters in rats and rabbits in conditions of oxidative stress caused by the administration of hydrogen peroxide. The SOD-CHS-CAT conjugate had antiplatelet potential due to its antiaggregation action manifested through the combination of enzyme activities and an acquired supramolecular structure. The influence on arterial fragment tonus was equivalent for SOD and CAT in native and conjugated form. Blood pressure and heart rate were significant and effectively normalized with SOD-CHS-CAT conjugate in rats and rabbits (after hydrogen peroxide administration as a perturbance stimulus). We have discovered the possibility of using the antioxidant bienzyme conjugate in chronic prophylaxis. It is important for a real development of the oral form of the SOD-CHS-CAT conjugate. These results indicate that the development of enzyme conjugates can be medically significant, as a promising approach for the creation of new drugs.
Maksimenko, A.V.; Vavaev, A.V.; Bouryachkovskaya, L.I.; Mokh, V.P.; Uchitel, I.A.; Lakomkin, V.L.; Kapelko, V.I.; Tischenko, E.G.
Study on the interaction of catalase with pesticides by flow injection chemiluminescence and molecular docking.
The interaction mechanisms of catalase (CAT) with pesticides (including organophosphates: disulfoton, isofenphos-methyl, malathion, isocarbophos, dimethoate, dipterex, methamidophos and acephate; carbamates: carbaryl and methomyl; pyrethroids: fenvalerate and deltamethrin) were first investigated by flow injection (FI) chemiluminescence (CL) analysis and molecular docking. By homemade FI-CL model of lg[(I0-I)/I]=lgK+nlg[D], it was found that the binding processes of pesticides to CAT were spontaneous with the apparent binding constants K of 10(3)-10(5)Lmol(-1) and the numbers of binding sites about 1.0. The binding abilities of pesticides to CAT followed the order: fenvalerate>deltamethrin>disulfoton>isofenphos-methyl>carbaryl>malathion>isocarbophos>dimethoate>dipterex>acephate>methomyl>methamidophos, which was generally similar to the order of determination sensitivity of pesticides. The thermodynamic parameters revealed that CAT bound with hydrophobic pesticides by hydrophobic interaction force, and with hydrophilic pesticides by hydrogen bond and van der Waals force. The pesticides to CAT molecular docking study showed that pesticides could enter into the cavity locating among the four subdomains of CAT, giving the specific amino acid residues and hydrogen bonds involved in CAT-pesticides interaction. It was also found that the lgK values of pesticides to CAT increased regularly with increasing lgP, Mr, MR and MV, suggesting that the hydrophobicity and steric property of pesticide played essential roles in its binding to CAT. PMID:24875908
Tan, Xijuan; Wang, Zhuming; Chen, Donghua; Luo, Kai; Xiong, Xunyu; Song, Zhenghua
Involvement of catalase in the protective benefits of metformin in mice with oxidative liver injury.
Metformin is a commonly used anti-diabetic drug with AMP-activated protein kinase (AMPK)-dependent hypoglycemic activities. Recent studies have revealed its anti-inflammatory and anti-oxidative properties. In the present study, the anti-oxidative potential of metformin and its potential mechanisms were investigated in a mouse model with carbon tetrachloride (CCl4)-induced severe oxidative liver injury. Our results showed that treatment with metformin significantly attenuated CCl4-induced elevation of serum aminotransferases and hepatic histological abnormalities. The alleviated liver injury was associated with decreased hepatic contents of oxidized glutathione (GSSG) and malondialdehyde (MDA). In addition, metformin treatment dose-dependently enhanced the activities of catalase (CAT) and decreased CCl4-induced elevation of hepatic H2O2 levels, but it had no obvious effects on the protein level of CAT. We also found that metformin increased the level of phosphorylated AMP-activated protein kinase (AMPK), but treatment with AMPK activator AICAR had no obvious effects on CAT activity. A molecular docking analysis indicated that metformin might interact with CAT via hydrogen bonds. These data suggested that metformin effectively alleviated CCl4-induced oxidative liver injury in mice and these hepatoprotective effects might be associated with CAT. PMID:24717679
Dai, Jie; Liu, Mingwei; Ai, Qing; Lin, Ling; Wu, Kunwei; Deng, Xinyu; Jing, Yuping; Jia, Mengying; Wan, Jingyuan; Zhang, Li
Catalasic activity in fish liver: improvement of the UV to visible analytic method.
Antioxidative defenses and more especially catalasic activity (CAT) are studied in a large range of scientific research thematics. In environmental sciences, the problematic of oxidative stress is of great interest as pollutants can induce perturbations of redox homeostasis. Consequently, changes in antioxidative defenses levels in fish tissues and particularly in liver are used as potential biomarkers of pollution. In most studies, the CAT was assayed by following during 5 min the consumption of H2O2 in cytosolic buffered extracts at 240 nm (UV-method). This study proposed a development of this method in the visible, using permanganate and a 525-nm detection, which was more accurate, sensitive, and rapid. Moreover, the hepatic CAT of six different fish species [a cyclidae (Nimbochromis linni), 3 cyprinidae (Brachydanio rerio, Rutilus rutilus, Cyprinus carpio), an anguillidae (Anguilla anguilla), and a percidae (Perca fluviatilus)] was evaluated with the two protocols (UV- and KMnO4-method). The results but also the thermal optimum of the reaction and the interest of CAT as biomarker in ecotoxicology were discussed. PMID:23224832
Paris-Palacios, S�verine; Delahaut, Laurence; Carreras, Alexis; Thomas, Marielle; Biagianti-Risbourg, Sylvie
Analysis of Oxidative Stress Status, Catalase and Catechol-O-Methyltransferase Polymorphisms in Egyptian Vitiligo Patients
Vitiligo is the most common depigmentation disorder of the skin. Oxidative stress is implicated as one of the probable events involved in vitiligo pathogenesis possibly contributing to melanocyte destruction. Evidence indicates that certain genes including those involved in oxidative stress and melanin synthesis are crucial for development of vitiligo. This study evaluates the oxidative stress status, the role of catalase (CAT) and catechol-O-Methyltransferase (COMT) gene polymorphisms in the etiology of generalized vitiligo in Egyptians. Total antioxidant capacity (TAC) and malondialdehyde (MDA) levels as well as CAT exon 9 T/C and COMT 158 G/A polymorphisms were determined in 89 patients and 90 age and sex-matched controls. Our results showed significantly lower TAC along with higher MDA levels in vitiligo patients compared with controls. Meanwhile, genotype and allele distributions of CAT and COMT polymorphisms in cases were not significantly different from those of controls. Moreover, we found no association between both polymorphisms and vitiligo susceptibility. In conclusion, the enhanced oxidative stress with the lack of association between CAT and COMT polymorphisms and susceptibility to vitiligo in our patients suggest that mutations in other genes related to the oxidative pathway might contribute to the etiology of generalized vitiligo in Egyptian population.
Mehaney, Dina A.; Darwish, Hebatallah A.; Hegazy, Rehab A.; Nooh, Mohammed M.; Tawdy, Amira M.; Gawdat, Heba I.; El-Sawalhi, Maha M.
Combined superoxide dismutase/catalase mimetics alter fetal pulmonary arterial smooth muscle cell growth.
Reactive oxygen species (ROS) are known to play an important role in the proliferation and viability of vascular smooth muscle cells. We have shown previously that treatment of fetal pulmonary arterial smooth muscle cells (FPASMC) with concentrations of 25 microM and higher of EUK-134, a superoxide dismutase/catalase mimetic, decreased cell viability via the induction of apoptosis. Here we demonstrate a dose-dependent decrease in serum-induced FPASMC growth at lower doses of EUK-134. This was due to the attenuation of FPASMC proliferation rather than the induction of apoptosis. Moreover, we found that the inhibition of FPASMC proliferation was observed using EUK-134 at concentrations as low as 5 microM. This inhibition of proliferation correlated with a 31% decrease in superoxide levels, as estimated using the oxidation of dihydroethidium. Flow cytometry revealed an increase in FPASMC in G2 after 24 h of exposure to 10 microM EUK-134. This was associated with a twofold increase in levels of the cell-cycle regulatory protein p21. This, together with our previous data, suggests that ROS levels determine the rate of FPASMC proliferation and, when below a threshold level, trigger apoptosis. Titration of ROS with antioxidants may help to prevent, or reverse, the vascular remodeling manifest in many cardiovascular disease states. PMID:14713351
Wedgwood, Stephen; Black, Stephen M
PaCATB, a secreted catalase protecting Podospora anserina against exogenous oxidative stress
A differential mass spectrometry analysis of secreted proteins from juvenile and senescent Podospora anserina cultures revealed age-related differences in protein profiles. Among other proteins with decreased abundance in the secretome of senescent cultures a catalase, termed PaCATB, was identified. Genetic modulation of the abundance of PaCATB identified differential effects on the phenotype of the corresponding strains. Deletion of PaCatB resulted in decreased resistance, over-expression in increased resistance against hydrogen peroxide. While the lifespan of the genetically modified strains was found to be unaffected under standard growth conditions, increased exogenous hydrogen peroxide stress in the growth medium markedly reduced the lifespan of the PaCatB deletion strain but extended the lifespan of PaCatB over-expressors. Overall our data identify a component of the secretome of P. anserina as a new effective factor to cope with environmental stress, stress that under natural conditions is constantly applied on organisms and influences aging processes.
Zintel, Sandra; Bernhardt, Dominik; Rogowska-Wrzesinska, Adelina; Osiewacz, Heinz D.
Association of polymorphic markers of the catalase and superoxide dismutase genes with type 2 diabetes mellitus.
Our study aims at determining whether genetic polymorphisms of catalase (CAT 1167C/T) and superoxide dismutase (SOD +35 A/C) could be associated with type 2 diabetes mellitus (T2DM). The study was conducted on 105 Egyptian patients with T2DM and 115 control subjects. Genotypes were done by polymerase chain reaction-restriction fragment length polymorphism methods. Homeostatic model assessment of insulin resistance (HOMA-IR), CAT and SOD activities, glycated hemoglobin, and insulin and lipid profiles were assessed. CAT and SOD activities were significantly decreased in T2DM compared with the control subjects. T allele of CAT and C allele of SOD1 were significant risk factors for T2DM. No effects of CAT or SOD1 gene polymorphisms on glycated haemoglobin or on HOMA-IR were found. With regard to the enzymes activities, only +35 A/C of SOD1 was related to SOD activity. Genetic variants C1167T of CAT gene and +35 A/C of SOD1 gene has no role in insulin resistance in T2DM. PMID:22970972
Ghattas, Maivel H; Abo-Elmatty, Dina M
T Cell Responses to Mycobacterial Catalase-Peroxidase Profile a Pathogenic Antigen in Systemic Sarcoidosis1
Sarcoidosis is a systemic granulomatous disease associated with local epithelioid granulomas, CD4+ T cells, and Th1 cytokines. The tissue Ags that drive this granulomatous inflammation are uncertain. In this study, we used IFN-?-ELISPOT assays and flow cytometry to assess lung and blood T cell responses to the candidate pathogenic Ag, Mycobacterium tuberculosis catalase-perox-idase (mKatG) in patients with sarcoidosis from two centers. Despite differences in patient phenotypic, genetic, and prognostic characteristics, we report that T cell responses to mKatG were remarkably similar in these cohorts, with higher frequencies of mKatG-reactive, IFN-?-expressing T cells in the blood of sarcoidosis patients compared with nontuberculosis sensitized healthy controls, and (in a subset) in greater numbers than T cells reactive to purified protein derivative. In sarcoidosis, mKatG-reactive CD4+ Th1 cells preferentially accumulated in the lung, indicating a compartmentalized response. Patients with or without L�fgren syndrome had similar frequencies of mKatG specific IFN-?-expressing blood T cells. Circulating mKatG-reactive T cells were found in chronic active sarcoidosis but not in patients with inactive disease. Together, these results demonstrate that T cell responses to mKatG in sarcoidosis fit a profile expected for a pathogenic Ag, supporting an immunotherapeutic approach to this disease.
Chen, Edward S.; Wahlstrom, Jan; Song, Zhimin; Willett, Matthew H.; Wiken, Maria; Yung, Rex C.; West, Erin E.; McDyer, John F.; Zhang, Ying; Eklund, Anders; Grunewald, Johan; Moller, David R.
Potential toxicity of sarafloxacin to catalase: Spectroscopic, ITC and molecular docking descriptions
NASA Astrophysics Data System (ADS)
The interaction between sarafloxacin and catalase (CAT) was studied by fluorescence spectroscopy, UV-visible absorption spectroscopy, circular dichroism (CD) spectroscopy, isothermal titration microcalorimetry (ITC) and molecular docking method. After deducting the inner filter effect, the fluorescence of CAT was quenched regularly by different concentrations of sarafloxacin. The quenching mechanism was studied by lifetime measurement, and it was proved to be mostly due to static quenching. The formation of sarafloxacin-CAT complex alters the micro-environment of amide moieties and tryptophan (Trp) residues, reduces the ?-helix content of the enzyme, changes the peripheral substituents on the porphyrin ring of heme and leads to the inhibition of the enzyme activity. Molecular docking study reveals that sarafloxacin is located between two ?-helix of CAT near to Trp 182 and Trp 185 residues, which supports the experimental results and helps to have a more clear understanding about the interaction mechanism. The change in the relative position of His 74 to heme induced by the variation of secondary structure is considered to be the major reason for the reduction of CAT activity. Moreover, sarafloxacin binds into a hydrophobic area of CAT mainly through hydrophobic interactions, which is consistent with the ITC analysis.
Cao, Zhaozhen; Liu, Rutao; Yang, Bingjun
A simple method to measure effective catalase activities: optimization, validation, and application in green coffee.
Oxidative metabolism in coffee cherries during maturation appears to be regulated by the timely expression of redox enzymes such as catalase (CAT), peroxidase (POD), and polyphenoloxidase (PPO). Among these enzymes, CAT is suspected to contribute significantly in setting the redox status of the healthy cherry and the processed bean. The initial redox status of the green bean might further control the nature and dynamics of reactions induced by roasting and eventually quality aspects of the end product. In this respect, Arabica (Coffea arabica) and Robusta (Coffea canephora) typically differ by their cup coffee flavor profiles. We developed an assay that allowed us to screen numerous green coffee samples for effective CAT activities. The proposed assay, which monitors CAT activities by online oxygen sensing in green coffee crude suspensions incubated with H2O2, seeks to integrate potential effects of endogenous inhibitors and activators. After optimization and validation of the assay, 23 Arabicas, 23 Robustas, and 8 Arabustas were analyzed. Nearly all Arabicas (22 of 23) harbored high CAT activity levels, whereas all Robustas harbored low ones. Arabustas performed like Arabicas of the lower CAT activity range. The traditional spectrophotometric assay did not reveal these specificities. Because of its simplicity, our assay might be valuable for assessing effective CAT activities in various plant tissues. PMID:17141173
Montavon, Philippe; Kukic, Koraljka Rade; Bortlik, Karlheinz
Influence of catalase gene silencing on the survivability of Sitobion avenae.
Reactive oxygen species (ROS), such as superoxide anions and hydrogen peroxide produced in cell metabolism, result in the disruption of cellular function and structure. Catalase (CAT), an enzyme which exists in almost all organisms including plants, invertebrates and vertebrates, acts in scavenging ROS. In this study, a sequence fragment encoding a CAT-like protein from wheat aphids (?Sitobion avenae) was cloned. Amino acid sequence alignment showed this CAT shared relatively high conservation with CAT sequences from other insects. We detected cat mRNA levels at nymphs of different stages and adults and results showed that cat expression in adults was significantly higher compared to juvenile stages. At the third instar stage, ingestion of dsCAT significantly knocked down CAT expression. Continuous feeding of dsCAT mixed in an artificial diet led to reduced survival rate and ecdysis index. This study indicates that cat, a potential target gene for management of insect pests, is important for maintaining the survival of ?S. avenae. PMID:24719312
Deng, Fei; Zhao, Zhangwu
Catalase-peroxidase activity is decreased in a Caulobacter crescentus rho mutant.
A Caulobacter crescentus rho::Tn5 mutant strain presenting a partially functional transcription termination factor Rho is highly sensitive to hydrogen peroxide in both exponential and stationary phases. The mutant was shown to be permanently under oxidative stress, based on fluorophore oxidation, and also to be sensitive to tert-butyl hydroperoxide and paraquat. However, the results showed that the activities of superoxide dismutases CuZnSOD and FeSOD and the alkylhydroperoxide reductase ahpC mRNA levels in the rho mutant were comparable to the wild-type control in the exponential and stationary phases. In contrast, the KatG catalase activity of the rho mutant strain was drastically decreased and did not show the expected increase in the stationary phase compared with the exponential phase. Transcription of the katG gene was increased in the rho mutant and the levels of the immunoreactive KatG protein do not differ considerably compared with the wild type in the stationary phase, suggesting that KatG activity is affected in a translational or a post-translational step. PMID:20002190
Italiani, Val�ria C S; Braz, V�nia S; Xiao, Huifang; Steinman, Howard M; Marques, Marilis V
Pancreatic adenocarcinoma up-regulated factor (PAUF), a novel up-regulated secretory protein in pancreatic ductal adenocarcinoma.
The identification of novel tumor-specific proteins or antigens is of great importance for diagnostic and therapeutic applications in pancreatic cancer. Using oligonucleotide microarrays, we identified a broad spectrum of differentially expressed pancreatic cancer-related genes. Of these, we selected an overexpressed expressed sequence taq and cloned a 721-bp full-length cDNA with an open reading frame of 196 amino acids. This novel gene was localized on the Homo sapiens 16p13.3 chromosomal locus, and its nucleotide sequence matched the Homo sapiens similar to common salivary protein 1 (LOC124220). We named the gene pancreatic adenocarcinoma up-regulated factor. The pancreatic adenocarcinoma up-regulated factor was secreted into the culture medium of pancreatic adenocarcinoma up-regulated factor-overexpressing Chinese hamster ovary cells, had an apparent molecular mass of approximately 25 kDa, and was N-glycosylated. The induction of pancreatic adenocarcinoma up-regulated factor in Chinese hamster ovary cells increased cell proliferation, migration, and invasion ability in vitro. Subcutaneous injection of mice with Chinese hamster ovary/pancreatic adenocarcinoma up-regulated factor cells resulted in 3.8-fold greater tumor sizes compared to Chinese hamster ovary/mock cells. Reverse transcription-polymerase chain reaction and western blotting with antirecombinant human pancreatic adenocarcinoma up-regulated factor antibodies confirmed that pancreatic adenocarcinoma up-regulated factor was highly expressed in six of eight pancreatic cancer cell lines. Immunohistochemical staining of human pancreatic cancer tissues also showed pancreatic adenocarcinoma up-regulated factor overexpression in the cytoplasm of cancer cells. Transfection with pancreatic adenocarcinoma up-regulated factor-specific small-interfering RNA reduced cancer cell migration and invasion in vitro. Treatment with antirecombinant human pancreatic adenocarcinoma up-regulated factor in vitro and in vivo reduced proliferation, migration, invasion, and tumorigenic ability. Collectively, our results suggest that pancreatic adenocarcinoma up-regulated factor is a novel secretory protein involved in pancreatic cancer progression and might be a potential target for the treatment of pancreatic cancer. PMID:19302292
Kim, Sun A; Lee, Yangsoon; Jung, Dawoon E; Park, Kyung Hwa; Park, Jeong Youp; Gang, Jingu; Jeon, Sun Bok; Park, Eui Chul; Kim, Young-Gun; Lee, Bogman; Liu, Qing; Zeng, Wen; Yeramilli, Subramanyam; Lee, Soojin; Koh, Sang Seok; Song, Si Young
Catalase expression in MCF-7 breast cancer cells is mainly controlled by PI3K/Akt/mTor signaling pathway.
Catalase is an antioxidant enzyme that catalyzes mainly the transformation of hydrogen peroxide into water and oxygen. Although catalase is frequently down-regulated in tumors the underlying mechanism remains unclear. Few transcription factors have been reported to directly bind the human catalase promoter. Among them FoxO3a has been proposed as a positive regulator of catalase expression. Therefore, we decided to study the role of the transcription factor FoxO3a and the phosphatidylinositol-3 kinase (PI3K) signaling pathway, which regulates FoxO3a, in the expression of catalase. To this end, we developed an experimental model of mammary breast MCF-7 cancer cells that acquire resistance to oxidative stress, the so-called Resox cells, in which catalase is overexpressed as compared with MCF-7 parental cell line. In Resox cells, Akt expression is decreased but its phosphorylation is enhanced when compared with MCF-7 cells. A similar profile is observed for FoxO3a, with less total protein but more phosphorylated FoxO3a in Resox cells, correlating with its higher Akt activity. The modulation of FoxO3a expression by knockdown and overexpression strategies did not affect catalase expression, neither in MCF-7 nor in Resox cells. Inhibition of PI3K and mTOR by LY295002 and rapamycin, respectively, decreases the phosphorylation of downstream targets (i.e. GSK3? and p70S6K) and leads to an increase of catalase expression only in MCF-7 but not in Resox cells. In conclusion, FoxO3a does not appear to play a critical role in the regulation of catalase expression in both cancer cells. Only MCF-7 cells are sensitive and dependent on PI3K/Akt/mTOR signaling. PMID:24630930
Glorieux, Christophe; Auquier, Julien; Dejeans, Nicolas; Sid, Brice; Demoulin, Jean-Baptiste; Bertrand, Luc; Verrax, Julien; Calderon, Pedro Buc
Dynamics of erythrocyte count, hemoglobin, and catalase activity in rat blood in hypokinesia, muscular activity and restoration
NASA Technical Reports Server (NTRS)
Tests were conducted to prove that muscular exertion (in this instance swimming) of different duration and intensity, as well as hypodynamia, result in an increase of hemoglobin and number of red blood cells in peripheral blood rats. Catalase activity increased with an increase in the duration of swimming, but only up to 6 hr; with 7-9 hr of swimming as well as in hypodynamia, catalase activity decreased. It was also observed that under hypodynamia as well as in 3, 5 and 6 hr exertion (swimming) the color index of blood decreased. Pressure chamber treatment (for 8 min each day for one week), alternating a 2 min negative pressure up to 35 mm Hg with 1 min positive pressure, increased the erythrocyte count and hemoglobin content.
Taneyeva, G. V.; Potapovich, G. M.; Voloshko, N. A.; Uteshev, A. B.
Crystallization and preliminary X-ray analysis of a bifunctional catalase-phenol oxidase from Scytalidium thermophilum
Catalase-phenol oxidase from Scytalidium thermophilum is a bifunctional enzyme: its major activity is the catalase-mediated decomposition of hydrogen peroxide, but it also catalyzes phenol oxidation. To understand the structural basis of this dual functionality, the enzyme, which has been shown to be a tetramer in solution, has been purified by anion-exchange and gel-filtration chromatography and has been crystallized using the hanging-drop vapour-diffusion technique. Streak-seeding was used to obtain larger crystals suitable for X-ray analysis. Diffraction data were collected to 2.8?� resolution at the Daresbury Synchrotron Radiation Source. The crystals belonged to space group P21 and contained one tetramer per asymmetric unit.
Sutay Kocabas, Didem; Pearson, Arwen R.; Phillips, Simon E. V.; Bakir, Ufuk; Ogel, Zumrut B.; McPherson, Michael J.; Trinh, Chi H.
Identification of lactic acid bacteria and Gram-positive catalase-positive cocci isolated from naturally fermented sausage (sucuk).
The aim of the study was to identify lactic acid bacteria and Gram-positive catalase-positive cocci isolated from Turkish dry fermented sausage (sucuk) produced by 7 different manufacturers without using starter culture. A total of 129 isolates of lactic acid bacteria were identified phenotypically. Lactobacillus plantarum was the dominant species (45.7%) followed by L. curvatus (10.9%) and L. fermentum (9.3%). Pediococcus isolates were identified as P. pentosaceus and P. acidilactici. All the isolates of gram-positive and catalase-positive cocci (123 isolates) were classified as Staphylococcus except for 1 isolate assigned to Kocuria rosea. The species isolated most often were S. xylosus (41.5%) and S. saprophyticus (28.5%). Four isolates were identified as S. equorum (3.3%), 1 isolate was assigned to S. carnosus (0.8%). PMID:19019118
Kaban, G; Kaya, M
Exogenous H 2O 2 and catalase treatments interfere with Tri genes expression in liquid cultures of Fusarium graminearum
Effect of exogenous H2O2 and catalase was tested in liquid cultures of the deoxynivalenol and 15-acetyldeoxynivalenol-producing fungus Fusarium graminearum. Accordingly to previous results, H2O2 supplementation of the culture medium leads to increased toxin production. This study indicates that this event seems to be linked to a general up regulation of genes involved in the deoxynivalenol and 15-acetyldeoxynivalenol biosynthesis pathway, commonly
Nadia Ponts; Laetitia Pinson-Gadais; Christian Barreau; Florence Richard-Forget; Th�r�se Ouellet
Cj1386 Is an Ankyrin-Containing Protein Involved in Heme Trafficking to Catalase in Campylobacter jejuni
Campylobacter jejuni, a microaerophilic bacterium, is the most frequent cause of human bacterial gastroenteritis. C. jejuni is exposed to harmful reactive oxygen species (ROS) produced during its own normal metabolic processes and during infection from the host immune system and from host intestinal microbiota. These ROS will damage DNA and proteins and cause peroxidation of lipids. Consequently, identifying ROS defense mechanisms is important for understanding how Campylobacter survives this environmental stress during infection. Construction of a ?Cj1386 isogenic deletion mutant and phenotypic assays led to its discovery as a novel oxidative stress defense gene. The ?Cj1386 mutant has an increased sensitivity toward hydrogen peroxide. The Cj1386 gene is located directly downstream from katA (catalase) in the C. jejuni genome. A ?katA? Cj1386 double deletion mutant was constructed and exhibited a sensitivity to hydrogen peroxide similar to that seen in the ?Cj1386 and ?katA single deletion mutants. This observation suggests that Cj1386 may be involved in the same detoxification pathway as catalase. Despite identical KatA abundances, catalase activity assays showed that the ?Cj1386 mutant had a reduced catalase activity relative to that of wild-type C. jejuni. Heme quantification of KatA protein from the ?Cj1386 mutant revealed a significant decrease in heme concentration. This indicates an important role for Cj1386 in heme trafficking to KatA within C. jejuni. Interestingly, the ?Cj1386 mutant had a reduced ability to colonize the ceca of chicks and was outcompeted by the wild-type strain for colonization of the gastrointestinal tract of neonate piglets. These results indicate an important role for Cj1386 in Campylobacter colonization and pathogenesis.
Flint, Annika; Sun, Yi-Qian
Cloning, expression and physiological analysis of broccoli catalase gene and Chinese cabbage ascorbate peroxidase gene under heat stress
The objectives of this work were to clone the catalase (CAT) gene from broccoli (Brassica oleracea) and the ascorbate peroxidase (APX) gene from Chinese cabbage and measure the regulation of CAT and APX gene expressions\\u000a under heat-stress conditions. Different genotypes responded differently to heat stress according to their various antioxidant\\u000a enzymes and physiological parameters. CAT and APX gene expression profiles
Kuan-Hung Lin; Ho-Chang Huang; Ching-Yun Lin
Effect of Nutrition Factors on the Synthesis of Superoxide Dismutase, Catalase, and Membrane Lipid Peroxide Levels in Cordyceps militaris Mycelium
Effect of carbon, nitrogen, and metal ion sources on superoxide dismutase (SOD), catalase (CAT) activities, and lipid perioxide\\u000a (LPO) levels in Cordyceps militaris mycelium were investigated at stationary growth phase by step supplementing with these nutrition factors in shake-flask cultures.\\u000a Mycelium was cultivated in several growth media containing different carbon sources. The observed highest SOD and CAT activities\\u000a were 44.3
Zun-sheng Wang; Yu-xiang Gu; Qin-sheng Yuan
Influence of different types of effectors on the kinetic parameters of suicide inactivation of catalase by hydrogen peroxide
The effects of cyanide and azide ions (class A), sodium-n-dodecyl sulphate (SDS) and 2-mercaptoethanol (class B), 3-aminotriazole (class C) and NADPH (class D) on the initial activity (ai), inactivation rate constant (ki) and the partition ratio (r) of bovine liver catalase reaction with its suicide substrate, hydrogen peroxide, were studied in 50 mM sodium phosphate buffer, pH 7.0 at 27�C.
Mahmood Ghadermarzi; Ali A Moosavi-Movahedi; Mohammad Ghadermarzi
Selective peracetic acid determination in the presence of hydrogen peroxide using a label free enzymatic method based on catalase
Peracetic acid (PAA) is selectively determined in the presence of hydrogen peroxide (H2O2) by using the self-indicating UV�Vis molecular absorption properties of catalase. The PAA reacts with the protein giving\\u000a an intermediate (Cat-I) which is reduced back by the aminoacid core surrounding the hemegroup. Since the original form of\\u000a the enzyme and the Cat-I have different UV�Vis absorption properties, the
Javier Galb�n; Vanesa Sanz; Susana de Marcos
Anti-apoptotic proteins and catalase-dependent apoptosis resistance in nickel chloride-transformed human lung epithelial cells
Chronic exposure to nickel compounds is associated with increased incidence of certain types of human cancer, including lung and nasal cancers. Despite intensive investigation, the oncogenic processes remain poorly understood. Apoptosis resistance is a key feature for tumor cells to escape physiological surveillance and acquire growth advantage over normal cells. Although NiCl2 exposure induces transformation of human lung epithelial cells, little information is available with regard to its molecular mechanisms, it is also not clear if the transformed cells are apoptosis resistant and tumorigenic. We explored the apoptosis resistance properties of nickel chloride-transformed human lung epithelial cells and the underlying mechanisms. The results showed that transformed BEAS-2B human lung epithelial cells are resistant to NiCl2-induced apoptosis. They have increased Bcl-2, Bcl-xL and catalase protein levels over the passage matched non-transformed counterparts. The mechanisms of apoptosis resistance are mitochondria-mediated and caspase-dependent. Forced overexpression of Bcl-2, Bcl-xL and catalase proteins reduced NiCl2-induced cell death; siRNA-mediated knockdown of their expression sensitized the cells to nickel-induced apoptosis, suggesting that Bcl-2, Bcl-xl and catalase protein expression plays a critical role in apoptosis resistance. Akt also participates in this process, as its overexpression increases Bcl-xL protein expression levels and attenuates NiCl2-induced apoptosis. Furthermore, transformed cells are tumorigenic in a xenograft model. Together, these results demonstrate that nickel-transformed cells are apoptosis-resistant and tumorigenic. Increased expression of Bcl-2, Bcl-xL and catalase proteins are important mechanisms contributing to transformed cell oncogenic properties.
Catalase in fluvial biofilms: a comparison between different extraction methods and example of application in a metal-polluted river
Antioxidant enzymes are involved in important processes of cell detoxification during oxidative stress and have, therefore,\\u000a been used as biomarkers in algae. Nevertheless, their limited use in fluvial biofilms may be due to the complexity of such\\u000a communities. Here, a comparison between different extraction methods was performed to obtain a reliable method for catalase\\u000a extraction from fluvial biofilms. Homogenization followed
Chlo� Bonnineau; Berta Bonet; Nat�lia Corcoll; Helena Guasch
Response of peroxidase and catalase to acid rain stress during seed germination of rice, wheat, and rape
Seed germination of plants with various acid-resistance display different responses to acid rain. To understand the reason\\u000a why such differences occur, the effects of simulated acid rain (pH 2.5�5.0) on the activities of peroxidase (POD) and catalase\\u000a (CAT) during seed germination of rice (O. sativa), wheat (T. aestivum), and rape (B. chinensis var. oleifera) were investigated. Results indicated that the
Lihong Wang; Xiaohua Huang; Qing Zhou
Biochemical biomarkers in environmental studies�lessons learnt from enzymes catalase, glutathione S -transferase and cholinesterase in two crustacean species
Background, aim and scope��For reliable environmental risk assessment of pollutants, knowledge on the effects at different levels of biological organisation\\u000a is needed. During the early days of biomarker research in environmental studies approximately two decades ago, biochemical\\u000a biomarkers were considered as the most promising tool for such purposes. Among these, three enzymes have often been studied:\\u000a catalase (CAT), glutathione S-transferase
Anita Jemec; Damjana Drobne; Tatjana Ti�ler; Kristina Sep?i?
Activation of galactose-containing glycoprotein and solid supports by galactose oxidase in presence of catalase for immobilization purposes
Specific oxidation of D-galactose present in the carbohydrate moiety of glucose oxidase from Aspergillus niger by galactose oxidase in the presence of catalase (48% efficiency) did not change the activity of the enzyme. Oxidized enzyme was coupled to hydrazide derivatives of O-a-D-galactosyl Separon H 1000 or of Sepharose 4B. Both solid supports were modified with adipic acid dihydrazide after their
L. Petkov; J. Sajdok; K. Rae; M. �?chov�; J. K�; J. Turkov�
Induction of apoptosis in fetal pulmonary arterial smooth muscle cells by a combined superoxide dismutase/catalase mimetic.
Reactive oxygen species (ROS) such as superoxide and hydrogen peroxide are known to play an important role in the proliferation and viability of vascular smooth muscle cells. In this study, we determined the effects of increased superoxide dismutase and catalase activity on fetal pulmonary arterial smooth muscle cell (FPASMC) proliferation and viability using EUK-134, a superoxide dismutase/catalase mimetic. Treatment of FPASMC with EUK-134 or with a combination of superoxide dismutase and catalase enzymes decreased superoxide and hydrogen peroxide levels as detected by the fluorescent dyes dihydroethidium and dichlorodihydrofluorescein diacetate, respectively. EUK-134 (5 microM) attenuated serum-induced FPASMC proliferation, whereas 50 microM EUK-134 decreased the number of viable cells, suggesting cell death. Conversely, combined superoxide dismutase and catalase enzyme activity equivalent to 50 microM EUK-134 prevented proliferation but did not reduce the number of viable FPASMC. The loss of mitochondrial membrane potential after 18 h, an increase in caspase-9 and caspase-3 activity after 24 h, and the subsequent appearance of TdT-mediated dUTP nick end labeling-positive nuclei were detected in FPASMC after treatment with 50 microM EUK-134. This indicates an induction of programmed rather than necrotic cell death and suggests that prolonged removal of ROS is required to stimulate apoptosis. Compounds such as EUK-134 may, therefore, prove more effective than enzymic antioxidants over longer periods, especially when the aim is to decrease the number of smooth muscle cells in diseases resulting from excessive muscularization. PMID:12665466
Wedgwood, Stephen; Black, Stephen M
High-performance liquid chromatography ultraviolet assay for human erythrocytic catalase activity by measuring glutathione as o-phthalaldehyde derivative
The most frequently used catalase (CAT) activity assay is based on the spectrophotometric measurement of hydrogen peroxide (H2O2) absorbance decrease at 240nm. Here we report an alternative high-performance liquid chromatography (HPLC) assay for human erythrocytic CAT (heCAT) activity measurement based on glutathione (GSH) analysis as a highly stable, H2O2-insensitive o-phthalaldehyde (OPA) derivative. The method was developed and validated using an
Anke B�hmer; Jens Jordan; Dimitrios Tsikas
A catalase-peroxidase from a newly isolated thermoalkaliphilic Bacillus sp. with potential for the treatment of textile bleaching effluents
A new thermoalkaliphilic bacterium was isolated from a textile wastewater drain and identified as a new Bacillus sp. (Bacillus SF). Because of its high pH stability and thermostability, a catalase-peroxidase (CP) from this strain has potential for the treatment of textile bleaching effluents. The CP from Bacillus SF was purified to more than 70.3-fold homogeneity using fractionated ammonium sulfate precipitation,
Marinka Gudelj; Gilbert Fruhwirth; Andreas Paar; Fritz Lottspeich; Karl-Heinz Robra; Artut Cavaco-Paulo; Georg G�bitz
Egg yolk peptides up-regulate glutathione synthesis and antioxidant enzyme activities in a porcine model of intestinal oxidative stress.
Long-term oxidative stress in the gastrointestinal tract can lead to the development of chronic intestinal disorders. Many food-derived antioxidants are effective in vitro, but the variable reports of in vivo efficacy and the pro-oxidant nature of some antioxidants necessitate alternative strategies for the reduction of in vivo oxidative stress. Compounds that up-regulate the production of endogenous antioxidants such as glutathione (GSH) and antioxidant enzymes provide novel approaches for the restoration of redox homeostatis. Egg yolk peptides (EYP) prepared from Alcalase and protease N digestion of delipidated egg yolk proteins were found to exhibit antioxidative stress properties. The effect of EYP supplementation was examined in a hydrogen peroxide-induced human colon cell line and in an animal model of intestinal oxidative stress. EYP significantly reduced the pro-inflammatory cytokine, IL-8, in Caco-2 cells. In piglets given intraperitoneal infusions of hydrogen peroxide, EYP treatment increased GSH and gamma-glutamylcysteine synthetase mRNA expression and activity, significantly increased antioxidant enzyme activities, in particular catalase and glutathione S-transferase activities, and reduced protein and lipid oxidation in the duodenum, jejunum, ileum, and colon. Furthermore, EYP boosted the systemic antioxidant status in blood by increasing the GSH concentration in red blood cells. These results suggest that EYP supplementation is a novel strategy for the reduction of intestinal oxidative stress. PMID:20540508
Young, Denise; Fan, Ming Z; Mine, Yoshinori
Gestational exposure to BDE-99 produces toxicity through upregulation of CYP isoforms and ROS production in the fetal rat liver.
On gestation day (GD) 6 to GD 19, pregnant Sprague Dawley rats were orally exposed to 0, 0.5, 1, and 2 mg/kg/day to one of the most prevalent polybrominated diphenyl ethers congeners found in humans, 2,2',4,4',5-pentaBDE (BDE-99). All dams were euthanized on GD 20, and live fetuses were evaluated for sex, body weight, and external, internal, and skeletal malformations and developmental variations. The liver from one fetus of each litter was excised for the evaluation of oxidative stress markers and the messenger RNA expression of multiple cytochrome P450 (CYP) isoforms. Exposure to BDE-99 during the gestational period produced delayed ossification, slight hypertrophy of the heart, and enlargement of the liver in fetuses. A transplacental effect of BDE-99, evidenced by the activation of nuclear hormones receptors that induce the upregulation of CYP1A1, CYP1A2, CYP2B1, and CYP3A2 isoforms, was also found in fetal liver. These isoforms are correlated with the activity level of the enzyme catalase and the levels of thiobarbituric acid reactive substances. However, teratogenic effects from BDE-99 exposure were not observed. Clear signs of embryo/fetal toxicity, due to a possible hormonal disruption, were evidenced by a large increase in the CYP system and the production of reactive oxygen species in fetal liver. PMID:22331496
Blanco, Jordi; Mulero, Miquel; Domingo, Jos� L; S�nchez, Dom�nec J
Up-Regulated Expression of AOS-LOXa and Increased Eicosanoid Synthesis in Response to Coral Wounding
In octocorals, a catalase�like allene oxide synthase (AOS) and an 8R-lipoxygenase (LOX) gene are fused together encoding for a single AOS-LOX fusion protein. Although the AOS-LOX pathway is central to the arachidonate metabolism in corals, its biological function in coral homeostasis is unclear. Using an acute incision wound model in the soft coral Capnella imbricata, we here test whether LOX pathway, similar to its role in plants, can contribute to the coral damage response and regeneration. Analysis of metabolites formed from exogenous arachidonate before and after fixed time intervals following wounding indicated a significant increase in AOS-LOX activity in response to mechanical injury. Two AOS-LOX isoforms, AOS-LOXa and AOS-LOXb, were cloned and expressed in bacterial expression system as active fusion proteins. Transcription levels of corresponding genes were measured in normal and stressed coral by qPCR. After wounding, AOS-LOXa was markedly up-regulated in both, the tissue adjacent to the incision and distal parts of a coral colony (with the maximum reached at 1 h and 6 h post wounding, respectively), while AOS-LOXb was stable. According to mRNA expression analysis, combined with detection of eicosanoid product formation for the first time, the AOS-LOX was identified as an early stress response gene which is induced by mechanical injury in coral.
Lohelaid, Helike; Teder, Tarvi; Toldsepp, Kadri; Ekins, Merrick; Samel, Nigulas
Manganese L-edge X-ray absorption spectroscopy of manganese catalase from Lactobacillus plantarum and mixed valence manganese complexes
The first Mn L-edge absorption spectra of a Mn metalloprotein are presented in this paper. Both reduced and superoxidized Mn catalase have been examined by fluorescence-detected soft X-ray absorption spectroscopy, and their Mn L-edge spectra are dramatically different. The spectrum of reduced Mn(II)Mn(II) catalase has been interpreted by ligand field atomic multiplet calculations and by comparison to model compound spectra. The analysis finds a 10 Dq value of nearly 1.1 eV, consistent with coordination by predominately nitrogen and oxygen donor ligands. For interpretation of mixed valence Mn spectra, an empirical simulation procedure based on the addition of homovalent model compound spectra has been developed and was tested on a variety of Mn complexes and superoxidized Mn catalase. This routine was also used to determine the oxidation state composition of the Mn in [Ba{sub 8}Na{sub 2}ClMn{sub 16}(OH){sub 8}(CO{sub 3}){sub 4}L{sub 8}] .53 H{sub 2}O (L=1,3-diamino-2-hydroxypropane-N,N,N`N`-tetraacetic acid). 27 refs., 6 figs.
Grush, M.M.; Chen, J.; George, S.J. [Univ. of California, Davis, CA (United States)] [and others] [Univ. of California, Davis, CA (United States); and others
The 2.2?� resolution structure of the catalase-peroxidase KatG from Synechococcus elongatus PCC7942.
The crystal structure of catalase-peroxidase from Synechococcus elongatus PCC7942 (SeKatG) was solved by molecular replacement and refined to an Rwork of 16.8% and an Rfree of 20.6% at 2.2?� resolution. The asymmetric unit consisted of only one subunit of the catalase-peroxidase molecule, including a protoporphyrin IX haem moiety and two sodium ions. A typical KatG covalent adduct was formed, Met248-Tyr222-Trp94, which is a key structural element for catalase activity. The crystallographic equivalent subunit was created by a twofold symmetry operation to form the functional dimer. The overall structure of the dimer was quite similar to other KatGs. One sodium ion was located close to the proximal Trp314. The location and configuration of the proximal cation site were very similar to those of typical peroxidases such as ascorbate peroxidase. These features may provide a structural basis for the behaviour of the radical localization/delocalization during the course of the enzymatic reaction. PMID:24598912
Kamachi, Saori; Wada, Kei; Tamoi, Masahiro; Shigeoka, Shigeru; Tada, Toshiji
Glutathione and catalase provide overlapping defenses for protection against respiration-generated hydrogen peroxide in Haemophilus influenzae.
Glutathione is an abundant and ubiquitous low-molecular-weight thiol that may play a role in many cellular processes, including protection against the deleterious effects of reactive oxygen species. We address here the role of glutathione in protection against hydrogen peroxide (H2O2) in Haemophilus influenzae and show that glutathione and catalase provide overlapping defense systems. H. influenzae is naturally glutathione deficient and imports glutathione from the growth medium. Mutant H. influenzae lacking catalase and cultured in glutathione-deficient minimal medium is completely devoid of H2O2 scavenging activity and, accordingly, substantial amounts of H2O2 accumulate in the growth medium. H. influenzae generates H2O2 at rates similar to those reported for Escherichia coli, but the toxicity of this harmful metabolite is averted by glutathione-based H2O2 removal, which appears to be the primary system for protection against H2O2 endogenously generated during aerobic respiration. When H2O2 concentrations exceed low micromolar levels, the hktE gene-encoded catalase becomes the predominant scavenger. The requirement for glutathione in protection against oxidative stress is analogous to that in higher and lower eukaryotes but is unlike the situation in other bacteria in which glutathione is dispensable for aerobic growth during both normal and oxidative stress conditions. PMID:12949108
Vergauwen, Bjorn; Pauwels, Frederik; Van Beeumen, Jozef J
Reactive oxygen species detoxification by catalase is a major determinant of fecundity in the mosquito Anopheles gambiae.
The mosquito Anopheles gambiae is a primary vector of Plasmodium parasites in Africa. The effect of aging on reproductive output in A. gambiae females from three strains that differ in their ability to melanize Plasmodium and in their systemic levels of hydrogen peroxide (H2O2), a reactive oxygen species (ROS), was analyzed. The number of eggs oviposited after the first blood meal decreases with age in all strains; however, this decline was much more pronounced in the G3 (unselected) and R (refractory to Plasmodium infection) strains than in the S (highly susceptible to Plasmodium) strain. Reduction of ROS levels in G3 and R females by administration of antioxidants reversed this age-related decline in fecundity. The S and G3 strains were fixed for two functionally different catalase alleles that differ at the second amino acid position (Ser2Trp). Biochemical analysis of recombinant proteins revealed that the Trp isoform has lower specific activity and higher Km than the Ser isoform, indicating that the former is a less efficient enzyme. The Trp-for-Ser substitution appears to destabilize the functional tetrameric form of the enzyme. Both alleles are present in the R strain, and Ser/Ser females had significantly higher fecundity than Trp/Trp females. Finally, a systemic reduction in catalase activity by dsRNA-mediated knockdown significantly reduced the reproductive output of mosquito females, indicating that catalase plays a central role in protecting the oocyte and early embryo from ROS damage. PMID:17284604
DeJong, Randall J; Miller, Lisa M; Molina-Cruz, Alvaro; Gupta, Lalita; Kumar, Sanjeev; Barillas-Mury, Carolina
Role of catalase in ethanol-induced conditioned taste aversion: a study with 3-amino-1,2,4-triazole.
Recent studies involved acetaldehyde, the first ethanol metabolite, in both the rewarding and aversive effects of ethanol consumption. Brain acetaldehyde is believed to originate mainly from local brain metabolism of ethanol by the enzyme catalase. Therefore, the inhibition of catalase by 3-amino-1,2,4-triazole (aminotriazole) may help to clarify the involvement of acetaldehyde in ethanol's hedonic effects. In the present study, multiple doses of both ethanol and aminotriazole were used to investigate the effects of catalase inhibition on ethanol-induced conditioned taste aversion (CTA). A separate microdialysis experiment investigated the effects of aminotriazole pretreatment on the time course of brain ethanol concentrations. Ethanol induced a dose-dependent CTA with a maximal effect after conditioning with 2.0 g/kg ethanol. Aminotriazole pretreatments dose-dependently potentiated the CTA induced by 1.0 g/kg ethanol. However, aminotriazole pretreatments did not alter the CTA induced by higher ethanol doses (1.5 and 2.0 g/kg) probably because a maximal aversion for saccharin was already obtained without aminotriazole. The results of the microdialysis experiment confirmed that the effects of aminotriazole cannot be attributed to local alterations of brain ethanol levels. The present study argues against a role for brain acetaldehyde in ethanol's aversive effects but in favor of its involvement in ethanol rewarding properties. PMID:12681527
Quertemont, Etienne; Escarabajal, M Dolores; De Witte, Philippe
Bioaccumulation of fullerene (C60 ) and corresponding catalase elevation in Lumbriculus variegatus.
Fullerene (C60 ), with its unique physical properties and nanometer size, has been mass-produced for many applications in recent decades. The increased likelihood of direct release into the environment has raised interest in understanding both the environmental fate and corresponding biological effects of fullerenes to living organisms. Because few studies have emphasized fullerene uptake and resulting biochemical responses by living organisms, a toxicity screening test and a 28-d bioaccumulation test for Lumbriculus variegatus were performed. No mortality was observed in the range of 0.05?mg?C60 /kg dry sediment to 11.33?mg?C60 /kg dry sediment. A biota-sediment accumulation factor of micron-sized fullerene agglomerates (�-C60 ) was 0.032?�?0.008 at day 28, which is relatively low compared with pyrene (1.62?�?0.22). Catalase (CAT) activity, an oxidative stress indicator, was elevated significantly on day 14 for L. variegatus exposed to �-C60 (p?=?0.034). This peak CAT activity corresponded to the highest body residues observed in the present study, 199?�?80?�g?C60 /kg dry weight sediment. Additionally, smaller C60 agglomerate size increased bioaccumulation potential in L. variegatus. The relationship between C60 body residue and the increased CAT activity followed a linear regression. All results suggest that C60 has a lower bioaccumulation potential than pyrene but a higher potential to induce oxidative stress in L. variegatus. Environ Toxicol Chem 2014;33:1135-1141. � 2014 SETAC. PMID:24477927
Wang, Jiafan; Wages, Mike; Yu, Shuangying; Maul, Jonathan D; Mayer, Greg; Hope-Weeks, Louisa; Cobb, George P
Protection from ethanol-induced limb malformations by the superoxide dismutase/catalase mimetic, EUK-134.
Based on previous in vitro studies that have illustrated prevention of ethanol-induced cell death by antioxidants, using an in vivo model, we have tested the anti-teratogenic potential of a potent synthetic superoxide dismutase plus catalase mimetic, EUK-134. The developing limb of C57BL/6J mice, which is sensitive to ethanol-induced reduction defects, served as the model system. On their ninth day of pregnancy, C57BL/6J mice were administered ethanol (two intraperitoneal doses of 2.9 g/kg given 4 h apart) alone or in combination with EUK-134 (two doses of 10 mg/kg). Pregnant control mice were similarly treated with either vehicle or EUK-134, alone. Within 15 h of the initial ethanol exposure, excessive apoptotic cell death was observed in the apical ectodermal ridge (AER) of the newly forming forelimb buds. Forelimb defects, including postaxial ectrodactyly, metacarpal, and ulnar deficiencies, occurred in 67.3% of the ethanol-exposed fetuses that were examined at 18 days of gestation. The right forelimbs were preferentially affected. No limb malformations were observed in control fetuses. Cell death in the AER of embryos concurrently exposed to ethanol and EUK-134 was notably reduced compared with that in embryos from ethanol-treated dams. Additionally, the antioxidant treatment reduced the incidence of forelimb malformations to 35.9%. This work illustrates that antioxidants can significantly improve the adverse developmental outcome that results from ethanol exposure in utero, diminishing the incidence and severity of major malformations that result from exposure to this important human teratogen. PMID:15208273
Chen, Shao-Yu; Dehart, Deborah B; Sulik, Kathleen K
Application of different molecular techniques for characterization of catalase-positive cocci isolated from sucuk.
This study was carried out for the characterization and discrimination of the indigenous Gram positive, catalase-positive cocci (GCC) population in sucuk, a traditional Turkish dry-fermented sausage. Sucuk samples, produced by the traditional method without starter culture were collected from 8 local producers in Kayseri/Turkey and a total of 116 GCC isolates were identified by using different molecular techniques. Two different molecular fingerprinting methods; namely, randomly amplified polymorphic DNA-PCR (RAPD-PCR) and repetitive extragenic palindrome-PCR (rep-PCR), were used for the clustering of isolates and identification at species level was carried out by full length sequencing of 16S rDNA. Combining the results obtained from molecular fingerprinting and 16S rDNA sequencing showed that the dominant GCC species isolated from the sucuk samples was Staphylococcus saprophyticus followed by Staphylococcus succinus and Staphylococcus equorum belonging to the Staphylococcus genus. Real-time PCR DNA melting curve analysis and high-resolution melting (HRM) analysis targeting the V1 + V3 regions of 16S rDNA were also applied for the discrimination of isolates belonging to different species. It was observed statistically different Tm values and species-specific HRM profiles for all except 2 species (S. saprophyticus and Staphylococcus xylosus) that have high 16S rDNA sequence similarity. The combination of rep-PCR and/or PCR-RAPD with 16S rRNA gene sequencing was an efficient approach for the characterization and identification of the GCC population in spontaneously fermented sucuk. On the other hand, intercalating dye assays were found to be a simple and very promising technique for the differentiation of the GCC population at species level. PMID:24410408
Kesmen, Z�lal; Yarimcam, Burcu; Aslan, Hakiye; Ozbekar, Esra; Yetim, Hasan
Enthalpy of Decomposition of Hydrogen Peroxide by Catalase at 25C (with Molar Extinction Coefficients of H2O2 Solutions in the UV).
National Technical Information Service (NTIS)
The enthalpy of decomposition of hydrogen peroxide by catalase has been determined calorimetrically in isotonic saline solutions at 25C. Extinction coefficients are also reported by hydrogen perioxide solutions in the ultraviolet. (Author)
D. P. Nelson L. A. Kiesow
Changes in catalase and glucose-6-phosphatase distribution patterns within oval cell compartment as possible differentiation markers during viral hepatocarcinogenesis in woodchucks
Cytochemical analysis at the ultrastructural level was performed to characterize expression of catalase and glucose-6-phosphatase (G6Pase) activity as possible differentiation markers in oval cells proliferating during hepatocarcinogenesis induced in woodchucks by chronic infection with the woodchuck hepatitis virus (WHV) and additional treatment with aflatoxin B1 (AFB1). Oval cells from WHV-carriers treated with AFB1 showed two types of catalase-positive organelles: 1)
Svetlana Radaeva; Peter Bannasch
Streptococcus didelphis sp. nov., a streptococcus with marked catalase activity isolated from opossums (Didelphis virginiana) with suppurative dermatitis and liver fibrosis
b-Haemolytic, catalase-positive, Gram-positive cocci that formed chains in broth media but did not react with Lancefield group antisera were isolated from skin lesions, spleen, liver and lungs of nine opossums, including eight from a research colony and one from a wildlife rehabilitation organization. The isolates had vigorous catalase activity that was retained on initial passage on non-blood-containing media, but this
Fred R. Rurangirwa; Charlene A. Teitzel; Jing Cui; Dorothy M. French; Patrick L. McDonough; Thomas Besser
How does the push\\/pull effect of the axial ligand influence the catalytic properties of Compound I of catalase and cytochrome P450?
Density functional theory (DFT) calculations on the chemoselective epoxidation versus hydroxylation reactions of propene by oxoiron porphyrin models mimicking the active sites of catalase, cytochrome P450 (P450) and horseradish peroxidase Compound I (CpdI) are presented. The catalase reactions are concerted and proceed via two-state reactivity patterns on competing doublet and quartet spin state surfaces, but the lowest barrier is the
Renyue Wang; Sam P. de Visser
Fate of the porphyrin cofactors during the light-dependent turnover of catalase and of the photosystem II reaction-center protein D1 in mature rye leaves
The apoprotein of the enzyme catalase (EC 220.127.116.11) was shown to exhibit a light-dependent turnover in leaves. Present results indicate that photoinactivation of the enzyme was not accompanied by a synchronous destruction and new synthesis of its heme moiety. In rye (Secale cereale L.) leaves the catalase content was not depleted in light when porphyrin synthesis was inhibited by gabaculine.
J. Feierabend; Silvia Dehne
Thermus thermophilus as a Cell Factory for the Production of a Thermophilic Mn-Dependent Catalase Which Fails To Be Synthesized in an Active Form in Escherichia coli
Thermostable Mn-dependent catalases are promising enzymes in biotechnological applications as H2O2-de- toxifying systems. We cloned the genes encoding Mn-dependent catalases from Thermus thermophilus HB27 and HB8 and a less thermostable mutant carrying two amino acid replacements (M129V and E293G). When the wild-type and mutant genes were overexpressed in Escherichia coli, unmodified or six-His-tagged proteins of the expected size were overproduced
Aurelio Hidalgo; Lorena Betancor; Renata Moreno; Olga Zafra; Felipe Cava; Roberto Fernandez-Lafuente; J. M. Guisan; J. Berenguer
Cardiac overexpression of catalase rescues cardiac contractile dysfunction induced by insulin resistance: role of oxidative stress, protein carbonyl formation and insulin sensitivity
Aims\\/hypothesis��Insulin resistance leads to oxidative stress and cardiac dysfunction. This study examined the impact of catalase on insulin-resistance-induced cardiac dysfunction, oxidative damage and insulin sensitivity.Methods��Insulin resistance was initiated in FVB and catalase-transgenic mice by 12�weeks of sucrose feeding. Contractile and intracellular Ca2+ properties were evaluated in cardiomyocytes including peak shortening (PS), time-to-PS (TPS), time-to-90% relengthening (TR90), half-width duration (HWD), maximal
F. Dong; C. X. Fang; X. Yang; X. Zhang; F. L. Lopez; J. Ren
Neuroprotective effect of MnTMPyP, a superoxide dismutase\\/catalase mimetic in global cerebral ischemia is mediated through reduction of oxidative stress and DNA fragmentation
Excessive generation of free radicals and decreased levels of the antioxidant enzymes such as superoxide dismutase (SOD) and catalase have been observed after brain ischemic reperfusion injury. In the present study, we have investigated the neuroprotective potential of MnTMPyP (Mn(III)tetrakis(1-methyl-4-pyridyl)porphyrin pentachloride), a SOD\\/Catalase mimetic in bilateral carotid artery occlusion model of global cerebral ischemia in Mongolian gerbils. Five minutes of
Shyam S. Sharma; Sangeetha Gupta
Purification and characterization of a catalase from the nonsulfur phototrophic bacterium Rhodobacter sphaeroides ATH 2.4.1 and its role in the oxidative stress response.
When challenged with reactive oxidants, the nonsulfur phototrophic bacterium Rhodobacter sphaeroides ATH 2.4.1 exhibited an oxidative stress response during both phototrophic and chemotrophic growth. Upon preincubation with 100 microM H2O2, catalase activity increased fivefold. Catalase was also induced by other forms of oxidative stress, heat-shock, ethanol treatment, and stationary-phase conditions. Only one band of catalase activity was detected after native and denaturing PAGE. The enzyme was purified 304-fold with a yield of 7%. The purified enzyme displayed a heterodimeric structure with subunits of 75 and 68 kDa, corresponding to a molecular mass of approximately 150 kDa for the native enzyme. The subunits had almost identical amino-terminal peptide sequences, sharing substantial similarity with other bacterial catalases. The enzyme exhibited an apparent Km of 40 mM and a Vmax of 285,000 U (mg protein)-1. Spectroscopic analysis indicated the presence of protoheme IX. The heme content calculated from pyridine hemochrome spectra was 0.43 mol per mol of enzyme. The enzyme had a broad pH optimum and was inhibited by cyanide, azide, hydroxylamine, 2-mercaptoethanol, and sodium dithionite. These data indicate that this catalase belongs to the class of monofunctional catalases. PMID:9575236
Terzenbach, D P; Blaut, M
Malondialdehyde and catalase as the serum biomarkers of allyl chloride-induced toxic neuropathy.
Chronic exposure to allyl chloride (AC) is known to produce a central-peripheral distal axonopathy. To access the biomarker of exposure and elucidate the mechanism of neuropathy induced by AC, we performed a longitudinal observational study of malondialdehyde (MDA), anti-reactive oxygen species (anti-ROS), glutathione (GSH), catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD) in rats serum and sciatic nerve after 0, 3, 6, 9 and 12 weeks of AC administration. AC was administrated to Wistar rats by gavage at a single dosage of 200 mg/kg/per dose (three times per week). Rats were sacrificed after 0, 3, 6, 9 and 12 weeks of AC treatment, serum and sciatic nerves were quickly collected at 4 degrees C. The results showed that MDA levels in serum (115.4 and 126.2%) and sciatic nerve (130.5 and 145.3%) significantly increased (p<0.05) on 3rd week of AC treatment and at gait score of 2, and further changes of MDA levels were observed after 6, 9 and 12 weeks and at gait score of 3 and 4. While a decrease (p<0.05) in the activities of CAT on 6th week of AC intoxication and at gait score of 2 was observed in serum (81.2 and 72.8%) and sciatic nerve (71.7 and 70.7%). The other antioxidants also decreased in serum and sciatic nerve after 3, 6 and 9, 12 weeks' intoxication and at gait score of 2, 3 and 4. Significant (p<0.05) positive correlations were observed between serum and sciatic nerve in MDA levels (r=0.9162 and 0.9551, respectively) and CAT (r=0.9410 and 0.9557, respectively) activities as time went on and symptoms developed. Thus, AC intoxication was associated with elevation of lipid peroxidation and reduction of antioxidative status, and the time dependent changes of these indexes in Wistar rats' serum and sciatic nerve occurred. The misbalance of lipid peroxidation and antioxidation status might be one of mechanisms of toxic neuropathy induced by AC. MDA and CAT could be served as the biomarkers of AC exposure to afford the early diagnosis of AC-induced toxic neuropathy. PMID:16938375
Wang, Qing-Shan; Zhang, Cui-Li; Zhao, Xiu-Lan; Yu, Su-Fang; Xie, Ke-Qin
Redox-sensitive up-regulation of eNOS by purple grape juice in endothelial cells: role of PI3-kinase/Akt, p38 MAPK, JNK, FoxO1 and FoxO3a.
The vascular protective effect of grape-derived polyphenols has been attributable, in part, to their direct action on blood vessels by stimulating the endothelial formation of nitric oxide (NO). The aim of the present study was to determine whether Concord grape juice (CGJ), which contains high levels of polyphenols, stimulates the expression of endothelial NO synthase (eNOS) in porcine coronary artery endothelial cells and, if so, to determine the signaling pathway involved. CGJ dose- and time-dependently increased eNOS mRNA and protein levels and this effect is associated with an increased formation of NO in endothelial cells. The stimulatory effect of CGJ on eNOS mRNA is not associated with an increased eNOS mRNA stability and inhibited by antioxidants such as MnTMPyP, PEG-catalase, and catalase, and by wortmannin (an inhibitor of PI3-kinase), SB 203580 (an inhibitor of p38 MAPK), and SP 600125 (an inhibitor of JNK). Moreover, CGJ induced the formation of reactive oxygen species (ROS) in endothelial cells and this effect is inhibited by MnTMPyP, PEG-catalase, and catalase. The CGJ-induced the phosphorylation of p38 MAPK and JNK kinases is abolished by MnTMPyP. CGJ induced phosphorylation of transcription factors FoxO1 and FoxO3a, which regulate negatively eNOS expression, and this effect is prevented by MnTMPyP, PEG-catalase, wortmannin, SB203580 and SP600125. Moreover, chromatin immunoprecipitation assay indicated that the FoxO3a protein is associated with the eNOS promoter in control cells and that CGJ induced its dissociation. Thus, the present study indicates that CGJ up-regulates the expression of eNOS mRNA and protein leading to an increased formation of NO in endothelial cells. The stimulatory effect of CGJ is a redox-sensitive event involving PI3-kinase/Akt, p38 MAPK and JNK pathways, and the inactivation of the FoxO transcription factors, FoxO1 and FoxO3a, thereby preventing their repression of the eNOS gene. PMID:23533577
Alhosin, Mahmoud; Anselm, Eric; Rashid, Sherzad; Kim, Jong Hun; Madeira, Socorro Vanesca Frota; Bronner, Christian; Schini-Kerth, Val�rie B
Redox-Sensitive Up-Regulation of eNOS by Purple Grape Juice in Endothelial Cells: Role of PI3-Kinase/Akt, p38 MAPK, JNK, FoxO1 and FoxO3a
The vascular protective effect of grape-derived polyphenols has been attributable, in part, to their direct action on blood vessels by stimulating the endothelial formation of nitric oxide (NO). The aim of the present study was to determine whether Concord grape juice (CGJ), which contains high levels of polyphenols, stimulates the expression of endothelial NO synthase (eNOS) in porcine coronary artery endothelial cells and, if so, to determine the signaling pathway involved. CGJ dose- and time-dependently increased eNOS mRNA and protein levels and this effect is associated with an increased formation of NO in endothelial cells. The stimulatory effect of CGJ on eNOS mRNA is not associated with an increased eNOS mRNA stability and inhibited by antioxidants such as MnTMPyP, PEG-catalase, and catalase, and by wortmannin (an inhibitor of PI3-kinase), SB 203580 (an inhibitor of p38 MAPK), and SP 600125 (an inhibitor of JNK). Moreover, CGJ induced the formation of reactive oxygen species (ROS) in endothelial cells and this effect is inhibited by MnTMPyP, PEG-catalase, and catalase. The CGJ-induced the phosphorylation of p38 MAPK and JNK kinases is abolished by MnTMPyP. CGJ induced phosphorylation of transcription factors FoxO1 and FoxO3a, which regulate negatively eNOS expression, and this effect is prevented by MnTMPyP, PEG-catalase, wortmannin, SB203580 and SP600125. Moreover, chromatin immunoprecipitation assay indicated that the FoxO3a protein is associated with the eNOS promoter in control cells and that CGJ induced its dissociation. Thus, the present study indicates that CGJ up-regulates the expression of eNOS mRNA and protein leading to an increased formation of NO in endothelial cells. The stimulatory effect of CGJ is a redox-sensitive event involving PI3-kinase/Akt, p38 MAPK and JNK pathways, and the inactivation of the FoxO transcription factors, FoxO1 and FoxO3a, thereby preventing their repression of the eNOS gene.
Rashid, Sherzad; Kim, Jong Hun; Frota Madeira, Socorro Vanesca; Bronner, Christian; Schini-Kerth, Valerie B.
Selective Cyclooxygenase2 Inhibition Upregulates Renal Cortical ?v Integrin Expression
Background: Cyclooxygenase-2 (COX-2), the inducible isoform of the cyclooxygenases, is upregulated in various inflammatory renal diseases and responsible for prostaglandin formation. As prostaglandins are known to influence cell adhesion processes, we investigated the effect of COX-2 inhibition on the expression of ?v integrins, which are also enhanced in renal diseases and control the adherence between the endothelium and the extracellular
Christoph Waldner; Gunhild Heise; Jutta Meyer-Kirchrath; Karsten Schr�r; Bernd Grabensee; Peter Heering
Cytochrome P450-type Hydroxylation and Epoxidation in a Tyrosine-liganded Hemoprotein, Catalase-related Allene Oxide Synthase*
The ability of hemoproteins to catalyze epoxidation or hydroxylation reactions is usually associated with a cysteine as the proximal ligand to the heme, as in cytochrome P450 or nitric oxide synthase. Catalase-related allene oxide synthase (cAOS) from the coral Plexaura homomalla, like catalase itself, has tyrosine as the proximal heme ligand. Its natural reaction is to convert 8R-hydroperoxy-eicosatetraenoic acid (8R-HPETE) to an allene epoxide, a reaction activated by the ferric heme, forming product via the FeIV�OH intermediate, Compound II. Here we oxidized cAOS to Compound I (FeV=O) using the oxygen donor iodosylbenzene and investigated the catalytic competence of the enzyme. 8R-hydroxyeicosatetraenoic acid (8R-HETE), the hydroxy analog of the natural substrate, normally unreactive with cAOS, was thereby epoxidized stereospecifically on the 9,10 double bond to form 8R-hydroxy-9R,10R-trans-epoxy-eicosa-5Z,11Z,14Z-trienoic acid as the predominant product; the turnover was 1/s using 100 ?m iodosylbenzene. The enantiomer, 8S-HETE, was epoxidized stereospecifically, although with less regiospecificity, and was hydroxylated on the 13- and 16-carbons. Arachidonic acid was converted to two major products, 8R-HETE and 8R,9S-eicosatrienoic acid (8R,9S-EET), plus other chiral monoepoxides and bis-allylic 10S-HETE. Linoleic acid was epoxidized, whereas stearic acid was not metabolized. We conclude that when cAOS is charged with an oxygen donor, it can act as a stereospecific monooxygenase. Our results indicate that in the tyrosine-liganded cAOS, a catalase-related hemoprotein in which a polyunsaturated fatty acid can enter the active site, the enzyme has the potential to mimic the activities of typical P450 epoxygenases and some capabilities of P450 hydroxylases.
Boeglin, William E.; Brash, Alan R.
Enantioselective oxidation of 2-hydroxy carboxylic acids by glycolate oxidase and catalase coexpressed in methylotrophic Pichia pastoris.
Glycolate oxidase (GO; (S)-2-hydroxyacid oxidase, EC 22.214.171.124) is a flavin mononucleotide (FMN)-dependent enzyme, which catalyzes the oxidation of 2-hydroxy carboxylic acids to the corresponding 2-keto acids. Catalase has been used as cocatalyst to decompose hydrogen peroxide produced in the reaction, thus limiting peroxide-based side reactions and GO deactivation. GO from spinach and catalase T from Saccharomyces cerevisiae previously coexpressed in Pichia pastoris strain NRRL Y-21001, was permeabilized and used for the oxidation of 3-phenyllactic acid, 3-indolelactic acid, 3-chlorolactic acid, 2-hydroxybutanoic acid, and 2-hydroxydecanoic acid to demonstrate high degree of selectivity to the (S)-enantiomers, leaving (R)-isomers intact. The rates of oxidation ranged from 1.3 to 120.0%, relative to the oxidation of lactic acid to pyruvic acid. The best substrates were 3-chlorolactic acid (110%) and 2-hydroxybutanoic acid (120%). Oxidation was carried out with (R)-, (S)-, and (RS)-3-phenyllactic acid, (RS)-lactic acid, and (RS)-2-hydroxybutanoic acid in 500 mL scale to characterize the products and stoichiometry of the reaction. All (RS)- and (S)-2-hydroxy acids produced 2-keto acids at close to the theoretical yield in 1-9 h. (R)-3-Phenyllactic acid was not oxidized over a period of 9 h. Addition of exogenous FMN and catalase were not required for this oxidation using double recombinant Pichia pastoris whole cells. As GO is absolutely specific to (S)-enantiomers, it can be used for resolution of racemic 2-hydroxy acids to (R)-2-hydroxy acids as well as for production of 2-keto acids. This is the first report on the selectivity of a broad range of 2-hydroxy acids by GO. PMID:20014430
Das, Shuvendu; Glenn, James H; Subramanian, Mani
Characterization of OxyR as a negative transcriptional regulator that represses catalase production in Corynebacterium diphtheriae.
Corynebacterium diphtheriae and Corynebacterium glutamicum each have one gene (cat) encoding catalase. In-frame ?cat mutants of C. diphtheriae and C. glutamicum were hyper-sensitive to growth inhibition and killing by H(2)O(2). In C. diphtheriae C7(?), both catalase activity and cat transcription decreased ~2-fold during transition from exponential growth to early stationary phase. Prototypic OxyR in Escherichia coli senses oxidative stress and it activates katG transcription and catalase production in response to H(2)O(2). In contrast, exposure of C. diphtheriae C7(?) to H(2)O(2) did not stimulate transcription of cat. OxyR from C. diphtheriae and C. glutamicum have 52% similarity with E. coli OxyR and contain homologs of the two cysteine residues involved in H(2)O(2) sensing by E. coli OxyR. In-frame ?oxyR deletion mutants of C. diphtheriae C7(?), C. diphtheriae NCTC13129, and C. glutamicum were much more resistant than their parental wild type strains to growth inhibition by H(2)O(2). In the C. diphtheriae C7(?) ?oxyR mutant, cat transcripts were about 8-fold more abundant and catalase activity was about 20-fold greater than in the C7(?) wild type strain. The oxyR gene from C. diphtheriae or C. glutamicum, but not from E. coli, complemented the defect in ?oxyR mutants of C. diphtheriae and C. glutamicum and decreased their H(2)O(2) resistance to the level of their parental strains. Gel-mobility shift, DNaseI footprint, and primer extension assays showed that purified OxyR from C. diphtheriae C7(?) bound, in the presence or absence of DTT, to a sequence in the cat promoter region that extends from nucleotide position -55 to -10 with respect to the +1 nucleotide in the cat ORF. These results demonstrate that OxyR from C. diphtheriae or C. glutamicum functions as a transcriptional repressor of the cat gene by a mechanism that is independent of oxidative stress induced by H(2)O(2). PMID:22438866
Kim, Ju-Sim; Holmes, Randall K
Reactive oxygen species formation and cell death in catalase-deficient tobacco leaf disks exposed to cadmium
The physiological responses of tobacco (Nicotiana tabacum L.) to oxidative stress induced by cadmium were examined with respect to reactive oxygen species (ROS) formation, antioxidant\\u000a enzymes activities, and cell death appearance in wild-type SR1 and catalase-deficient CAT1AS plants. Leaf disks treated with\\u000a 100 or 500��M CdCl2 increased Evans blue staining and leakage of electrolytes in SR1 or CAT1AS plants, more
Mar�a Florencia Iannone; Eliana Paola Rosales; Mar�a Daniela Groppa; Mar�a Patricia Benavides
Bacterioferritin A Modulates Catalase A (KatA) Activity and Resistance to Hydrogen Peroxide in Pseudomonas aeruginosa
We have cloned a 3.6-kb genomic DNA fragment from Pseudomonas aeruginosa harboring the rpoA, rplQ, katA, and bfrA genes. These loci are predicted to encode, respectively, (i) the ? subunit of RNA polymerase; (ii) the L17 ribosomal protein; (iii) the major catalase, KatA; and (iv) one of two iron storage proteins called bacterioferritin A (BfrA; cytochrome b1 or b557). Our goal was to determine the contributions of KatA and BfrA to the resistance of P. aeruginosa to hydrogen peroxide (H2O2). When provided on a multicopy plasmid, the P. aeruginosa katA gene complemented a catalase-deficient strain of Escherichia coli. The katA gene was found to contain two translational start codons encoding a heteromultimer of ?160 to 170 kDa and having an apparent Km for H2O2 of 44.7 mM. Isogenic katA and bfrA mutants were hypersusceptible to H2O2, while a katA bfrA double mutant demonstrated the greatest sensitivity. The katA and katA bfrA mutants possessed no detectable catalase activity. Interestingly, a bfrA mutant expressed only ?47% the KatA activity of wild-type organisms, despite possessing wild-type katA transcription and translation. Plasmids harboring bfrA genes encoding BfrA altered at critical amino acids essential for ferroxidase activity could not restore wild-type catalase activity in the bfrA mutant. RNase protection assays revealed that katA and bfrA are on different transcripts, the levels of which are increased by both iron and H2O2. Mass spectrometry analysis of whole cells revealed no significant difference in total cellular iron levels in the bfrA, katA, and katA bfrA mutants relative to wild-type bacteria. Our results suggest that P. aeruginosa BfrA may be required as one source of iron for the heme prosthetic group of KatA and thus for protection against H2O2.
Ma, Ju-Fang; Ochsner, Urs A.; Klotz, Martin G.; Nanayakkara, Vagira K.; Howell, Michael L.; Johnson, Zaiga; Posey, James E.; Vasil, Michael L.; Monaco, John J.; Hassett, Daniel J.
Bacterioferritin A modulates catalase A (KatA) activity and resistance to hydrogen peroxide in Pseudomonas aeruginosa.
We have cloned a 3.6-kb genomic DNA fragment from Pseudomonas aeruginosa harboring the rpoA, rplQ, katA, and bfrA genes. These loci are predicted to encode, respectively, (i) the alpha subunit of RNA polymerase; (ii) the L17 ribosomal protein; (iii) the major catalase, KatA; and (iv) one of two iron storage proteins called bacterioferritin A (BfrA; cytochrome b1 or b557). Our goal was to determine the contributions of KatA and BfrA to the resistance of P. aeruginosa to hydrogen peroxide (H2O2). When provided on a multicopy plasmid, the P. aeruginosa katA gene complemented a catalase-deficient strain of Escherichia coli. The katA gene was found to contain two translational start codons encoding a heteromultimer of approximately 160 to 170 kDa and having an apparent Km for H2O2 of 44.7 mM. Isogenic katA and bfrA mutants were hypersusceptible to H2O2, while a katA bfrA double mutant demonstrated the greatest sensitivity. The katA and katA bfrA mutants possessed no detectable catalase activity. Interestingly, a bfrA mutant expressed only approximately 47% the KatA activity of wild-type organisms, despite possessing wild-type katA transcription and translation. Plasmids harboring bfrA genes encoding BfrA altered at critical amino acids essential for ferroxidase activity could not restore wild-type catalase activity in the bfrA mutant. RNase protection assays revealed that katA and bfrA are on different transcripts, the levels of which are increased by both iron and H2O2. Mass spectrometry analysis of whole cells revealed no significant difference in total cellular iron levels in the bfrA, katA, and katA bfrA mutants relative to wild-type bacteria. Our results suggest that P. aeruginosa BfrA may be required as one source of iron for the heme prosthetic group of KatA and thus for protection against H2O2. PMID:10368148
Ma, J F; Ochsner, U A; Klotz, M G; Nanayakkara, V K; Howell, M L; Johnson, Z; Posey, J E; Vasil, M L; Monaco, J J; Hassett, D J
A novel thermo-alkali stable catalase�peroxidase from Oceanobacillus oncorhynchi subsp. incaldaniensis : purification and characterization
A novel thermo-alkali-stable catalase�peroxidase from Oceanobacillus oncorhynchi subsp. incaldaniensis subsp. nov., strain 20AG, was purified and characterized. The protein purified from the cells resulted in 110-fold purification\\u000a with a specific activity of 35,000�U\\/mg. The enzyme consisted of four identical subunits of 72�kDa as determined by SDS-PAGE\\u000a and the total molecular mass measured by gel filtration was 280�kDa. The heme content
Valeria Calandrelli; Agata Gambacorta; Ida Romano; Vito Carratore; Licia Lama
Catalase activity is stimulated by H2O2 in rich culture medium and is required for H2O2 resistance and adaptation in yeast?
Catalases are efficient scavengers of H2O2 and protect cells against H2O2 stress. Examination of the H2O2 stimulon in Saccharomyces cerevisiae revealed that the cytosolic catalase T (Ctt1) protein level increases 15-fold on H2O2 challenge in synthetic complete media although previous work revealed that deletion of the CCT1 or CTA1 genes (encoding peroxisomal/mitochondrial catalase A) does not increase the H2O2 sensitivity of yeast challenged in phosphate buffer (pH 7.4). This we attributed to our observation that catalase activity is depressed when yeast are challenged with H2O2 in nutrient-poor media. Hence, we performed a systematic comparison of catalase activity and cell viability of wild-type yeast and of the single catalase knockouts, ctt1? and cta1?, following H2O2 challenge in nutrient-rich medium (YPD) and in phosphate buffer (pH 7.4). Ctt1 but not Cta1 activity is strongly induced by H2O2 when cells are challenged in YPD but suppressed when cells are challenged in buffer. Consistent with the activity results, exponentially growing ctt1? cells in YPD are more sensitive to H2O2 than wild-type or cta1? cells, whereas in buffer all three strains exhibit comparable H2O2 hypersensitivity. Furthermore, catalase activity is increased during adaptation to sublethal H2O2 concentrations in YPD but not in buffer. We conclude that induction of cytosolic Ctt1 activity is vital in protecting yeast against exogenous H2O2 but this activity is inhibited by H2O2 when cells are challenged in nutrient-free media.
Martins, Dorival; English, Ann M.
Mechanism of catalase activity in aqueous solutions of dimanganese(III,IV) ethylenediamine-N,N prime -diacetate
Manganous ions, ligated by ethylenediamine-N,N{prime}-diacetate (edda = L) decompose hydrogen peroxide with a rate law {minus}d(H{sub 2}O{sub 2})/dt = k{sub 17}(Mn(edda))(H{sub 2}O{sub 2}) where k{sub 17} = 5.4 M{sup {minus}1} s{sup {minus}1} at pH 7. The reduction of peroxide to water is initiated by the reaction of Mn{sup II}L with a dinuclear Mn{sup III,IV}L{sub 2}. A subsequent fast reaction between the transient product of this reaction and hydrogen peroxide or tert-butyl hydroperoxide effectively oxidizes Mn(II) to Mn(IV) in a concerted step without formation of the hydroxyl radical. The green mixed-valence complex, which is probably a bis({mu}-oxo)-bridged structure, is stable in neutral aqueous solution and exhibits a 16-line ESR signal in frozen solution. The basis of catalase activity is the autocatalytic formation of this complex when hydrogen peroxide is reduced by manganese(II). The catalase cycle is independent of the formation of oxy radicals. Mononuclear Mn{sup III}edda and Mn{sup II}edda react with superoxide radicals, but the decomposition of peroxide is virtually independent of these reactions. In unbuffered solutions, with a moderate excess of hydrogen peroxide, an oscillation in the concentration of the dinuclear complex is detected. 28 refs., 10 figs., 5 tabs.
Rush, J.D.; Maskos, Z. (Louisiana State Univ., Baton Rouge (USA))
A genetic approach to study H2O2 scavenging in fission yeast--distinct roles of peroxiredoxin and catalase.
The main peroxiredoxin in Schizosaccharomyces pombe, Tpx1, is important to sustain aerobic growth, and cells lacking this protein are only able to grow on solid plates under anaerobic conditions. We have found that deletion of the gene coding for thioredoxin reductase, trr1, is a suppressor of the sensitivity to aerobic growth of ?tpx1 cells, so that cells lacking both proteins are able to grow on solid plates in the presence of oxygen. We have investigated this suppression effect, and determined that it depends on the presence of catalase, which is constitutively expressed in ?trr1 cells in a transcription factor Pap1-dependent manner. A complete characterization of the repertoire of hydrogen peroxide scavenging activities in fission yeast suggests that Tpx1 is the only enzyme with sufficient sensitivity for peroxides and cellular abundance as to control the low levels produced during aerobic growth, catalase being the next barrier of detoxification when the steady-state levels of peroxides are increased in ?tpx1 cells. Gpx1, the only glutathione peroxidase encoded by the S.?pombe genome, only has a minor secondary role when extracellular peroxides are added. Our study proposes non-overlapping roles for the different hydrogen peroxide scavenging activities of this eukaryotic organism. PMID:24521463
Paulo, Esther; Garc�a-Santamarina, Sarela; Calvo, Isabel A; Carmona, Merc�; Boronat, Susanna; Dom�nech, Alba; Ayt�, Jos�; Hidalgo, Elena
Engineering of chimeric catalase-Angiopep-2 for intracellular protection of brain endothelial cells against oxidative stress.
Blood-brain barrier (BBB) disruption and brain microvascular endothelial cells (BMVECs) death caused by excessive production of hydrogen peroxide (H2O2) have been implicated in several neurological conditions. To overcome this problem, H2O2-degrading enzyme with ability to enter the BMVECs is required. In the present study, genetic fusion of gene encoding human catalase and gene encoding Angiopep-2 (AP2), a brain targeting peptide, was performed. The fusion protein was successfully expressed in Escherichia coli and purified to homogeneity. The protein retained heme content and specific enzymatic activity in the same order of magnitude as that of native enzyme. Study of the BMVECs internalization showed that 0.1?M of the fusion protein can enter the cell within 15min, while internalization of the native protein was not observed at this condition. In addition, treatment of the BMVECs with 20 units of the fusion protein for 30min showed protection against H2O2 up to 5.0mM, whereas this protective effect was not observed from treatment with the native protein. Therefore, construction of chimeric human catalase and AP2 provides an insight into the development of potential therapeutic antioxidant with ability to penetrate the BBB for protection against neurodegenerative disorders. PMID:24769213
Yainoy, Sakda; Houbloyfa, Patcharaporn; Eiamphungporn, Warawan; Isarankura-Na-Ayudhya, Chartchalerm; Prachayasittikul, Virapong
The effect of some factors of polluted environment on catalase responses and resistance of microbial isolates against toxic oxidative stress.
The properties of bacterial isolates from polluted environments which are characterized by increased levels of oxidative stress do not reflect only the level of contaminants, but also arise as a consequence of many permanently changed conditions. The survival rate of Comamonas terrigena N3H isolates from an environment with elevated levels of H(2)O(2) is correlated with stimulation of catalase. The response of bacterial catalase to the effect of phenol in exogenous conditions was affected by the presence of an additional contaminant, Cd(2+). An isolate of Aspergillus niger selected from river sediment containing 363�mg/kg As, 93�mg/kg Sb at pH 5.2-4.8 grew on Czapek-Dox agar ~1.6 times faster than an isolate of the same species from coal dust sediment with approximately the same level of pollution (400�mg/kg As) but somewhat lower pH (3.3-2.8). It also exhibited differences in the microscopic characteristics of its mycelial structures. Both isolates exhibited a higher tolerance to the exogenic toxic effects of metals (As(5+), Cd(2+), and Cu(2+) at 5, 25, or 50�mg/L) than a control culture, but the differences in tolerance between them were only slight. These laboratory results suggest that there are complicated relationships which may exist in the "in situ" environment. PMID:22706798
Polek, Bystr�k; Godo?�kov�, Jana
KatP, a novel catalase-peroxidase encoded by the large plasmid of enterohaemorrhagic Escherichia coli O157:H7.
A gene coding for a catalase-peroxidase activity was identified on a 9-7 kb Smal DNA fragment derived from the large plasmid pO157 of enterohaemorrhagic Escherichia coli (EHEC) O157:H7 strain EDL 933. Nucleotide sequencing revealed an ORF of 2208 bp and predicted a 736 amino acid polypeptide with a molecular mass of 81.8 kDa. This putative protein was found to be highly homologous to members of the bacterial bifunctional catalase-peroxidase family. Analysis of its amino acid sequence revealed the presence of characteristic peroxidase 1 and 2 motifs. In addition, an N-terminal signal sequence was found, suggesting that the catalase-peroxidase is transported through the cytoplasmic membrane. EHEC catalase-peroxidase activities were investigated in cytoplasmic and periplasmic crude extracts as well as in culture supernatants from wild-type and recombinant E. coli strains. EHEC-specific catalase-peroxidase activity was detected primarily in the periplasm in strain EDL 933. The newly discovered enzyme was designated KatP, to indicate its plasmid origin. PCR analysis of representative strains of all enteric E. coli pathogroups (i.e. enterohaemorrhagic, enterotoxigenic, enteropathogenic, enteroaggregative and enteroinvasive E. coli) revealed a close association between the occurrence of EHEC-haemolysin and the katP gene in Shiga-like-toxin-producing E. coli O157 strains. PMID:8969527
Brunder, W; Schmidt, H; Karch, H
Aspergillus nidulans Catalase-Peroxidase Gene (cpeA) Is Transcriptionally Induced during Sexual Development through the Transcription Factor StuA
Catalases, peroxidases, and catalase-peroxidases are important enzymes to cope with reactive oxygen species in pro- and eukaryotic cells. In the filamentous fungus Aspergillus nidulans three monofunctional catalases have been described, and a fourth catalase activity was observed in native polyacrylamide gels. The latter activity is probably due to the bifunctional enzyme catalase-peroxidase, which we characterized here. The gene, named cpeA, encodes an 81-kDa polypeptide with a conserved motif for heme coordination. The enzyme comprises of two similar domains, suggesting gene duplication and fusion during evolution. The first 439 amino acids share 22% identical residues with the C terminus. Homologous proteins are found in several prokaryotes, such as Escherichia coli and Mycobacterium tuberculosis (both with 61% identity). In fungi the enzyme has been noted in Penicillium simplicissimum, Septoria tritici, and Neurospora crassa (69% identical amino acids) but is absent from Saccharomyces cerevisiae. Expression analysis in A. nidulans revealed that the gene is transcriptionally induced upon carbon starvation and during sexual development, but starvation is not sufficient to reach high levels of the transcript during development. Besides transcriptional activation, we present evidence for posttranscriptional regulation. A green fluorescent protein fusion protein localized to the cytoplasm of H�lle cells. The H�lle cell-specific expression was dependent on the developmental regulator StuA, suggesting an activating function of this helix-loop-helix transcription factor.
Scherer, Mario; Wei, Huijun; Liese, Ralf; Fischer, Reinhard
HDAC inhibition upregulates the expression of angiostatic ADAMTS1.
HDAC inhibitors are promising anticancer agents that induce cell cycle arrest and apoptosis. However, the role of HDACs in cancer progression, such as angiogenesis and metastasis, remains largely unexplored. Among various HDAC inhibitors, we demonstrate that TSA and SAHA upregulated the expression of angiostatic ADAMTS1 in A549 cells. HDAC6 inhibitor tubacin, and knockdown of HDAC6, also lead to ADAMTS1 upregulation. By reporter, DAPA, and ChIP assays, the proximal GC boxes were demonstrated to be essential for ADAMTS1 induction. Decreased binding of SP1 and HDAC6 to the ADAMTS1 promoter after TSA treatment was also seen. These data suggest the involvement of HDAC6 and SP1 in the HDACi-induced expression of angiostatic ADAMTS1. PMID:19007777
Chou, Chia-Wei; Chen, Ching-Chow
Tetherin upregulation in simian immunodeficiency virus-infected macaques.
Here we show that simian immunodeficiency virus (SIV) infection of rhesus macaques results in rapid upregulation of tetherin (BST-2 or CD317) on peripheral blood lymphocytes, including the CD4(+) CCR5(+) T cell targets of virus infection, with a peak of induction that coincides with peak alpha interferon (IFN-?) levels in plasma, and that tetherin remains above baseline levels throughout chronic infection. These observations are consistent with a role for tetherin in innate immunity to immunodeficiency virus infection. PMID:24109219
Rahmberg, Andrew R; Neidermyer, William J; Breed, Matthew W; Alvarez, Xavier; Midkiff, Cecily C; Piatak, Michael; Lifson, Jeffrey D; Evans, David T
Tetherin Upregulation in Simian Immunodeficiency Virus-Infected Macaques
Here we show that simian immunodeficiency virus (SIV) infection of rhesus macaques results in rapid upregulation of tetherin (BST-2 or CD317) on peripheral blood lymphocytes, including the CD4+ CCR5+ T cell targets of virus infection, with a peak of induction that coincides with peak alpha interferon (IFN-?) levels in plasma, and that tetherin remains above baseline levels throughout chronic infection. These observations are consistent with a role for tetherin in innate immunity to immunodeficiency virus infection.
Rahmberg, Andrew R.; Neidermyer, William J.; Breed, Matthew W.; Alvarez, Xavier; Midkiff, Cecily C.; Piatak, Michael; Lifson, Jeffrey D.
Upregulated Genes In Sporadic, Idiopathic Pulmonary Arterial Hypertension
Background To elucidate further the pathogenesis of sporadic, idiopathic pulmonary arterial hypertension (IPAH) and identify potential therapeutic avenues, differential gene expression in IPAH was examined by suppression subtractive hybridisation (SSH). Methods Peripheral lung samples were obtained immediately after removal from patients undergoing lung transplant for IPAH without familial disease, and control tissues consisted of similarly sampled pieces of donor lungs not utilised during transplantation. Pools of lung mRNA from IPAH cases containing plexiform lesions and normal donor lungs were used to generate the tester and driver cDNA libraries, respectively. A subtracted IPAH cDNA library was made by SSH. Clones isolated from this subtracted library were examined for up regulated expression in IPAH using dot blot arrays of positive colony PCR products using both pooled cDNA libraries as probes. Clones verified as being upregulated were sequenced. For two genes the increase in expression was verified by northern blotting and data analysed using Student's unpaired two-tailed t-test. Results We present preliminary findings concerning candidate genes upregulated in IPAH. Twenty-seven upregulated genes were identified out of 192 clones examined. Upregulation in individual cases of IPAH was shown by northern blot for tissue inhibitor of metalloproteinase-3 and decorin (P < 0.01) compared with the housekeeping gene glyceraldehydes-3-phosphate dehydrogenase. Conclusion Four of the up regulated genes, magic roundabout, hevin, thrombomodulin and sucrose non-fermenting protein-related kinase-1 are expressed specifically by endothelial cells and one, muscleblind-1, by muscle cells, suggesting that they may be associated with plexiform lesions and hypertrophic arterial wall remodelling, respectively.
Edgar, Alasdair J; Chacon, Matilde R; Bishop, Anne E; Yacoub, Magdi H; Polak, Julia M
Uromodulin Upregulates TRPV5 by Impairing Caveolin-Mediated Endocytosis
Uromodulin (UMOD) is synthesized in the thick ascending limb and secreted into urine as the most abundant protein. Association studies in humans suggest protective effects of UMOD against calcium-containing kidney stones. Mice carrying mutations of Umod found in human uromodulin-associated kidney disease (UAKD) and Umod deficient mice exhibit hypercalciuria. The mechanism for UMOD regulation of urinary Ca2+ excretion is incompletely understood. We examined if UMOD regulates TRPV5 and TRPV6, channels critical for renal transcellular Ca2+ reabsorption. Coexpression with UMOD increased whole-cell TRPV5 current density in HEK293 cells. In biotinylation studies UMOD increased TRPV5 cell-surface abundance. Extracellular application of purified UMOD upregulated TRPV5 current density within physiological relevant concentration ranges. UMOD exerted a similar effect on TRPV6. TRPV5 undergoes constitutive caveolin-mediated endocytosis. UMOD had no effect on TRPV5 in a caveolin-1 deficient cell line. Expression of recombinant caveolin-1 in these cells restored the ability of UMOD to upregulate TRPV5. Secretion of UAKD-mutant UMOD was markedly reduced and coexpression of mutant UMOD with TRPV5 failed to increase its current. Immunofluorescent studies demonstrated lower TRPV5 expression in Umod?/? mice compared to wild-type. UMOD upregulates TRPV5 by acting from extracellular and by decreasing endocytosis of TRPV5. The stimulation of Ca2+ reabsorption via TRPV5 by UMOD may contribute to protection against kidney stone formation.
Wolf, Matthias T.F.; Wu, Xue-Ru; Huang, Chou-Long
Effects of the kinematic viscosity and surface tension on the bubble take-off period in a catalase-hydrogen peroxide system.
The effect of kinematic viscosity and surface tension of the solution was investigated by adding catalase, glucose oxidase, or glucose on the bubble movement in a catalase-hydrogen peroxide system. The kinematic viscosity was measured using a Cannon-Fenske kinematic viscometer. The surface tension of the solution was measured by the Wilhelmy method using a self-made apparatus. The effects of the hole diameter/cell wall thickness, catalase concentration, glucose concentration, and glucose oxidase concentration on the kinematic viscosity, surface tension, and bubble take-off period were investigated. With our system, the effects of the changes in the solution materiality on the bubble take-off period were proven to be very small in comparison to the change in the oxygen-producing rate. PMID:19250805
Sasaki, Satoshi; Iida, Yoshinori
Piper betle shows antioxidant activities, inhibits MCF-7 cell proliferation and increases activities of catalase and superoxide dismutase
Background Breast cancer is the most common form of cancer and the focus on finding chemotherapeutic agents have recently shifted to natural products. Piper betle is a medicinal plant with various biological activities. However, not much data is available on the anti-cancer effects of P. betle on breast cancer. Due to the current interest in the potential effects of antioxidants from natural products in breast cancer treatment, we investigated the antioxidant activities of the leaves of P. betle and its inhibitory effect on the proliferation of the breast cancer cell line, MCF-7. Methods The leaves of P. betle were extracted with solvents of varying polarities (water, methanol, ethyl acetate and hexane) and their phenolic and flavonoid content were determined using colorimetric assays. Phenolic composition was characterized using HPLC. Antioxidant activities were measured using FRAP, DPPH, superoxide anion, nitric oxide and hyroxyl radical scavenging assays. Biological activities of the extracts were analysed using MTT assay and antioxidant enzyme (catalase, superoxide dismutase, glutathione peroxidase) assays in MCF-7 cells. Results Overall, the ethyl acetate extract showed the highest ferric reducing activity and radical scavenging activities against DPPH, superoxide anion and nitric oxide radicals. This extract also contained the highest phenolic content implying the potential contribution of phenolics towards the antioxidant activities. HPLC analyses revealed the presence of catechin, morin and quercetin in the leaves. The ethyl acetate extract also showed the highest inhibitory effect against the proliferation of MCF-7 cells (IC50=65 ?g/ml). Treatment of MCF-7 cells with the plant extract increased activities of catalase and superoxide dismutase. Conclusions Ethyl acetate is the optimal solvent for the extraction of compounds with antioxidant and anti-proliferative activities. The increased activities of catalase and superoxide dismutase in the treated cells could alter the antioxidant defense system, potentially contributing towards the anti-proliferative effect. There is great potential for the ethyl acetate extract of P. betle leaf as a source of natural antioxidants and to be developed as therapeutics in cancer treatment.
Ascorbate peroxidase and catalase cooperate for protection against hydrogen peroxide generated in potato tubers during low-temperature storage.
We investigated the behavior of the antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APx), in potato tubers during storage at low temperature. SOD activity increased temporarily within 3 weeks and was higher at 1 degree C than at 20 degrees C. APx activity also increased more at low (1 degree C) than at higher temperatures (5 and 20 degrees C). The contents of ascorbic acid (AsA), which is the substrate of APx, decreased immediately within 3 weeks and then gradually decreased until 15 weeks. The activity of CAT, the other enzyme which can scavenge hydrogen peroxide, decreased once in the first six weeks and thereafter increased to 15 weeks. Thus, the enhancement of the active oxygen-scavenging system that was induced by low temperature in potato tubers could result not only in a decrease of AsA but also in combined increases in APx and CAT activity whose manners were different. PMID:9584985
Mizuno, M; Kamei, M; Tsuchida, H
Polymer-Induced Heteronucleation for Protein Single Crystal Growth: Structural Elucidation of Bovine Liver Catalase and Concanavalin A Forms
Obtaining single crystals for X-ray diffraction remains a major bottleneck in structural biology; when existing crystal growth methods fail to yield suitable crystals, often the target rather than the crystallization approach is reconsidered. Here we demonstrate that polymer-induced heteronucleation, a powerful technique that has been used for small molecule crystallization form discovery, can be applied to protein crystallization by optimizing the heteronucleant composition and crystallization formats for crystallizing a wide range of protein targets. Applying these advances to two benchmark proteins resulted in dramatically increased crystal size, enabling structure determination, for a half century old form of bovine liver catalase (BLC) that had previously only been characterized by electron microscopy, and the discovery of two new forms of concanavalin A (conA) from the Jack bean and accompanying structural elucidation of one of these forms.
Foroughi, Leila M.; Kang, You-Na; Matzger, Adam J. (Michigan)
Vitamin A supplementation induces oxidative stress and decreases the immunocontent of catalase and superoxide dismutase in rat lungs.
Lungs require an adequate supply of vitamin A for normal embryonic development, postnatal maturation, and maintenance and repair during adult life. However, recent intervention studies revealed that supplementation with retinoids resulted in a higher incidence of lung cancer, although the mechanisms underlying this effect are still unknown. Here, the authors studied the effect of vitamin A supplementation on oxidative stress parameters in lungs of Wistar rats. Vitamin A supplementation either at therapeutic (1000 and 2500 IU/kg) or excessive (4500 and 9000 IU/kg) doses for 28 days induced lipid peroxidation, protein carbonylation, and oxidation of protein thiol groups, as well as change in catalase (EC 188.8.131.52; CAT) and superoxide dismutase (EC 18.104.22.168, SOD) activities and immunocontents. These results altogether suggest that vitamin A supplementation causes significant changes in redox balance the free radical status in lungs, which are frequently associated to severe lung dysfunction. PMID:19842843
Pasquali, Matheus A B; Gelain, Daniel P; Oliveira, Marcos R; Behr, Guilherme A; Motta, Leonardo L; Rocha, Ricardo F; Klamt, F�bio; Moreira, Jos� C F
Synthetic superoxide dismutase/catalase mimetics reduce oxidative stress and prolong survival in a mouse amyotrophic lateral sclerosis model.
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder that causes motoneuron degeneration, paralysis and death. Mutations in Cu, Zn superoxide dismutase (SOD1) are one cause of this disease. It is widely suspected that increased reactive oxidative species (ROS) is involved in motoneuron degeneration but whether such an involvement plays a role in ALS progression in vivo is uncertain. We treated mice expressing human mutant SOD1 G93A with EUK-8 and EUK-134, two synthetic SOD/catalase mimetics that have shown efficacy in several animal models of human diseases. These treatments reduced levels of oxidative stress and prolonged survival. The results suggest that oxidative stress plays an active role in ALS and illustrate the potential for treatment strategies aimed specifically against ROS. PMID:11343826
Jung, C; Rong, Y; Doctrow, S; Baudry, M; Malfroy, B; Xu, Z
Lifespan extension and rescue of spongiform encephalopathy in superoxide dismutase 2 nullizygous mice treated with superoxide dismutase-catalase mimetics.
Superoxide is produced as a result of normal energy metabolism within the mitochondria and is scavenged by the mitochondrial form of superoxide dismutase (sod2). Mice with inactivated SOD2 (sod2 nullizygous mice) die prematurely, exhibiting several metabolic and mitochondrial defects and severe tissue pathologies, including a lethal spongiform neurodegenerative disorder (Li et al., 1995; Melov et al., 1998, 1999). We show that treatment of sod2 nullizygous mice with synthetic superoxide dismutase (SOD)-catalase mimetics extends their lifespan by threefold, rescues the spongiform encephalopathy, and attenuates mitochondrial defects. This class of antioxidant compounds has been shown previously to extend lifespan in the nematode Caenorhabditis elegans (Melov et al., 2000). These new findings in mice suggest novel therapeutic approaches to neurodegenerative diseases associated with oxidative stress such as Friedreich ataxia, spongiform encephalopathies, and Alzheimer's and Parkinson's diseases, in which chronic oxidative damage to the brain has been implicated. PMID:11606622
Melov, S; Doctrow, S R; Schneider, J A; Haberson, J; Patel, M; Coskun, P E; Huffman, K; Wallace, D C; Malfroy, B
[Effect of lipolytic and catalase activity on physico-mechanical properties of coating Polyken 980-25].
Lipolytic and catalase activity of Pseudomonas pseudoalcaligenes 109, Rhodococcus erythropolis 102, Bacillus subtilis 138 and their association with different growth models: biofilm and plankton ones. It is shown that under biofilm conditions the fermentative activity of bacteria under study was 1.5-1.7 times higher than under plankton conditions. Monocultures of bacteria displayed much lower activity than associative ones. Changes of physico-chemical properties of the specimens of protective coating Polyken 980-25 with participation of the above bacteria have been studied. The coating breaking strength decreases by 5.9-11.8% under the effect of monocultures, and by 17.3% under the effect of association. The adhesive strength as the basic index of coating biologic resistance decreased, respectively, in mono- and associated cultures by 28.6-73.2% in respect of the control. Damaging the sticking layer of isolation coating, bacteria damage the adhesion to metal which favors its corrosion. PMID:23516839
Kopteva, Zh P; Zanina, V V; Boretskaia, M A; Kopteva, A E; Kozlova, I A
Effect of N+ Beam Exposure on Superoxide Dismutase and Catalase Activities and Induction of Mn-SOD in Deinococcus Radiodurans
NASA Astrophysics Data System (ADS)
Though bacteria of the radiation-resistant Deinococcus radiodurans have a high resistance to the lethal and mutagenic effects of many DNA-damaging agents, the mechanisms involved in the response of these bacteria to oxidative stress are poorly understood. In this report, the superoxide dismutase (SOD) and catalase (CAT) activities produced by these bacteria were measured, and the change of SOD and CAT activities by 20 keV N+ beam exposure was examined. Their activities were increased by N+ beam exposure from 8�1014 ions/cm2 to 6�1015 ions/cm2. The treatment of H2O2 and [CHCl3 +CH3CH2OH] and the measurement of absorption spectrum showed that the increase in SOD activity was resulted from inducible activities of Mn-SOD in D. radiodurans AS1.633 by N+ beam exposure. These results suggested that this bacteria possess inducible defense mechanisms against the deleterious effects of oxidization.
Song, Dao-jun; Chen, Ruo-lei; Shao, Chun-lin; Wu, Li-jun; Yu, Zeng-liang
A study of the relative importance of the peroxiredoxin-, catalase-, and glutathione-dependent systems in neural peroxide metabolism.
Cells are endowed with several overlapping peroxide-degrading systems whose relative importance is a matter of debate. In this study, three different sources of neural cells (rat hippocampal slices, rat C6 glioma cells, and mouse N2a neuroblastoma cells) were used as models to understand the relative contributions of individual peroxide-degrading systems. After a pretreatment (30 min) with specific inhibitors, each system was challenged with either H?O? or cumene hydroperoxide (CuOOH), both at 100 ?M. Hippocampal slices, C6 cells, and N2a cells showed a decrease in the H?O? decomposition rate (23-28%) by a pretreatment with the catalase inhibitor aminotriazole. The inhibition of glutathione reductase (GR) by BCNU (1,3-bis(2-chloroethyl)-1-nitrosourea) significantly decreased H?O? and CuOOH decomposition rates (31-77%). Inhibition of catalase was not as effective as BCNU at decreasing cell viability (MTT assay) and cell permeability or at increasing DNA damage (comet test). Impairing the thioredoxin (Trx)-dependent peroxiredoxin (Prx) recycling by thioredoxin reductase (TrxR) inhibition with auranofin neither potentiated peroxide toxicity nor decreased the peroxide-decomposition rate. The results indicate that neural peroxidatic systems depending on Trx/TrxR for recycling are not as important as those depending on GSH/GR. Dimer formation, which leads to Prx2 inactivation, was observed in hippocampal slices and N2a cells treated with H?O?, but not in C6 cells. However, Prx-SO? formation, another form of Prx inactivation, was observed in all neural cell types tested, indicating that redox-mediated signaling pathways can be modulated in neural cells. These differences in Prx2 dimerization suggest specific redox regulation mechanisms in glia-derived (C6) compared to neuron-derived (N2a) cells and hippocampal slices. PMID:21440059
Mitozo, P�ricles Arruda; de Souza, Luiz Felipe; Loch-Neckel, Gecioni; Flesch, Samira; Maris, Angelica Francesca; Figueiredo, Cl�udia Pinto; Dos Santos, Adair Roberto Soares; Farina, Marcelo; Dafre, Alcir Luiz
Mycobacterial catalase-peroxidase is a tissue antigen and target of the adaptive immune response in systemic sarcoidosis
Sarcoidosis is a disease of unknown etiology characterized by noncaseating epithelioid granulomas, oligoclonal CD4+ T cell infiltrates, and immune complex formation. To identify pathogenic antigens relevant to immune-mediated granulomatous inflammation in sarcoidosis, we used a limited proteomics approach to detect tissue antigens that were poorly soluble in neutral detergent and resistant to protease digestion, consistent with the known biochemical properties of granuloma-inducing sarcoidosis tissue extracts. Tissue antigens with these characteristics were detected with immunoglobulin (Ig)G or F(ab?)2 fragments from the sera of sarcoidosis patients in 9 of 12 (75%) sarcoidosis tissues (150�160, 80, or 60�64 kD) but only 3 of 22 (14%) control tissues (all 62�64 kD; P = 0.0006). Matrix-assisted laser desorption/ionization time of flight mass spectrometry identified Mycobacterium tuberculosis catalase�peroxidase (mKatG) as one of these tissue antigens. Protein immunoblotting using anti-mKatG monoclonal antibodies independently confirmed the presence of mKatG in 5 of 9 (55%) sarcoidosis tissues but in none of 14 control tissues (P = 0.0037). IgG antibodies to recombinant mKatG were detected in the sera of 12 of 25 (48%) sarcoidosis patients compared with 0 of 11 (0%) purified protein derivative (PPD)? (P = 0.0059) and 4 of 10 (40%) PPD+ (P = 0.7233) control subjects, suggesting that remnant mycobacterial catalase�peroxidase is one target of the adaptive immune response driving granulomatous inflammation in sarcoidosis.
Song, Zhimin; Marzilli, Lisa; Greenlee, Brian M.; Chen, Edward S.; Silver, Richard F.; Askin, Frederic B.; Teirstein, Alvin S.; Zhang, Ying; Cotter, Robert J.; Moller, David R.
Oxygen-Dependent Regulation of the Expression of the Catalase Gene katA of Lactobacillus sakei LTH677
The catalase gene katA of Lactobacillus sakei LTH677 was cloned and expressed in Escherichia coli UM2, Lactobacillus casei LK1, and Lactobacillus curvatus LTH1432. The last host is a catalase-deficient plasmid-cured derivative of a starter organism used in meat fermentation. The regulation of katA expression was found to be the same in L. sakei LTH677 and the recombinant strains. The addition of H2O2 to anaerobic cultures, as well as a switch to aerobic conditions, resulted in a strong increase in KatA activity. The expression was investigated in more detail with L. sakei LTH677 and L. curvatus LTH4002. The recombinant strain LTH4002 did not accumulate H2O2 under glucose-limited aerobic conditions and remained viable in the stationary phase. Under inductive conditions, the katA-specific mRNA and the apoenzyme were synthesized de novo. Deletion derivatives of the katA promoter were produced, and the regulatory response was investigated by fusion to the ?-glucuronidase reporter gene gusA and expression in L. sakei LTH677. The fact that gene expression was subject to induction was confirmed at the level of transcription and protein synthesis. A small putative regulatory sequence of at least 25 bp was identified located upstream of the ?35 site. Competition experiments performed with L. sakei LTH677 harboring the fusion constructs consisting of the katA promoter and gusA revealed that an activator protein is involved in the transcriptional induction of katA.
Hertel, Christian; Schmidt, Gudrun; Fischer, Marc; Oellers, Katja; Hammes, Walter P.
Molecular, technological and safety characterization of Gram-positive catalase-positive cocci from slightly fermented sausages.
The population of Gram-positive catalase-positive cocci from slightly fermented sausages was characterized at species and strain level by molecular techniques and some technological and hygienic aspects were also considered. Staphylococcus xylosus was the predominant species (80.8%) followed by Staphylococcus warneri (8.3%), Staphylococcus epidermidis (5.8%) Staphylococcus carnosus (4.6%), and Kocuria varians (0.4%). Proteolytic activity was observed in 23% of the isolates. The species with the highest percentage of proteolytic strains was S. warneri. Lipolytic activity was found in 45.8% of the isolates and S. xylosus was the species with the highest percentage of lipolytic isolates. Biogenic amine production was not widely distributed (only 14.6% of the isolates). Tyramine was the most intense amine produced, although by only 4.6% of the isolates. Phenylethylamine was more frequently detected (10.8% of isolates) but at lower levels. Some strains also produced putrescine (3.3%), cadaverine (2.9%), histamine (1.3%) and tryptamine (0.4%). All isolates were susceptible to linezolid and vancomicin and over 70% were resistant to penicillin G, ampicillin and sulphonamides. Most of the mecA+ strains (only 4.6% of isolates) also displayed resistance to multiple antibiotics. A reduced enterotoxigenic potential was found. Only 3.3% of isolates showed staphylococcal enterotoxins genes, all identified as entC gene. The combination of RAPD-PCR and plasmid profiling allowed the discrimination of 208 different profiles among the 240 Gram-positive catalase-positive cocci characterized, indicating a great genetic variability. PMID:16297478
Mart�n, B; Garriga, M; Hugas, M; Bover-Cid, S; Veciana-Nogu�s, M T; Aymerich, T
Up-Regulated Dicer Expression in Patients with Cutaneous Melanoma
Background MicroRNAs (miRNAs) are small non-coding RNAs (18�24 nucleotides) that have recently been shown to regulate gene expression during cancer progression. Dicer, a central enzyme in the multi-component miRNA biogenesis pathway, is involved in cutting precursor miRNAs to functionally mature forms. Emerging evidence shows that Dicer expression is deregulated in some human malignancies and it correlates with tumor progression, yet this role has not yet been investigated in skin cancers. Methods and Findings Using an anti-human monoclonal antibody against Dicer and immunohistochemistry, we compared the expression of Dicer protein among 404 clinically annotated controls and skin tumors consisting of melanocytic nevi (n?=?71), a variety of melanomas (n?=?223), carcinomas (n?=?73) and sarcomas (n?=?12). Results showed a cell-specific up-regulated Dicer in 81% of cutaneous, 80% of acrolentiginous and 96% of metastatic melanoma specimens compared to carcinoma or sarcoma specimens (P<0.0001). The expression of Dicer was significantly higher in melanomas compared to benign melanocytic nevi (P<0.0001). In patients with cutaneous melanomas, Dicer up-regulation was found to be significantly associated with an increased tumor mitotic index (P?=?0.04), Breslow's depth of invasion (P?=?0.03), nodal metastasis (P?=?0.04) and a higher American Joint Committee on Caner (AJCC) clinical stage (P?=?0.009). Using western blot analysis, we confirmed the cell-specific up-regulation of Dicer protein in vitro. A pooled-analysis on mRNA profiling in cutaneous tumors showed up-regulation of Dicer at the RNA level in cutaneous melanoma, also showing deregulation of other enzymes that participate in the biogenesis and maturation of canonical miRNAs. Conclusions Increased Dicer expression may be a clinically useful biomarker for patients with cutaneous melanoma. Understanding deregulation of Dicer and its influence on miRNA maturation is needed to predict the susceptibility of melanoma patients to miRNA-based therapy in the future.
Ma, Zhihai; Swede, Helen; Cassarino, David; Fleming, Elizabeth; Fire, Andrew; Dadras, Soheil S.
Molecular Characterization of a Catalase-Negative Staphylococcus aureus subsp. aureus Strain Collected from a Patient with Mitral Valve Endocarditis and Pericarditis Revealed a Novel Nonsense Mutation in the katA Gene ?
We report a case of endocarditis and pericarditis caused by catalase-negative Staphylococcus aureus. Molecular characterization revealed a novel nonsense mutation in the katA gene, leading to a loss of 238 amino acids (47% of the wild-type catalase protein), including the heme-binding site, NADPH-binding region, and Tyr-337, essential for catalysis.
To, Kelvin K. W.; Cheng, Vincent C. C.; Chan, Jasper F. W.; Wong, Amy C. Y.; Chau, Sandy; Tsang, Flora H. F.; Curreem, Shirly O. T.; Lau, Susanna K. P.; Yuen, Kwok-Yung; Woo, Patrick C. Y.
Copper and Zinc-containing Superoxide Dismutase, Manganese-containing Superoxide Dismutase, Catalase, and Glutathione Peroxidase in Normal and Neoplastic Human Cell Lines and Normal Human Tissues1
Copper- and zinc-containing Superoxide dismutase, man ganese-containing Superoxide dismutase, catalase, and gluta- thione peroxidase form the primary enzymic defense against toxic oxygen reduction metabolites. Such metabolites have been implicated in the damage brought about by ionizing radiation, as well as in the effects of several cytostatic com pounds. These enzymes were analyzed in 31 different human normal diploid and neoplastic
Stefan L. Marklund; N. Gunnar Westman; Erik Lundgren; Goran Roos
Reduced susceptibility to waterlogging together with high-light stress is related to increases in superoxide dismutase and catalase activities in sweet potato
We investigated the changes in antioxidative enzyme activities of two sweet potato cultivars under waterlogging and high-light conditions in the growth chamber. The activities of antioxidative enzymes were measured from leaf crude extract of sweet potato during the first five days of the treatments. Activities of superoxide dismutase and catalase were consistently increased in Taoyuan 1 sweet potato over time
Shih-Ying Hwang; Hui-Wen Lin; Ruey-Houng Chern; Hsiao-Feng Lo; Liang Li
The role played by acid and basic centers in the activity of biomimetic catalysts of the catalase, peroxidase, and monooxidase reactions
The acid-basic centers of heterogeneous carriers of catalase, peroxidase, and monooxigenase biomimetics, in particular, iron protoporphyrin deposited on active or neutral aluminum magnesium silicate, were studied. The catalytic activity of biomimetics was stabilized, which allowed us not only to synthesize fairly effective biomimetics but also to clarify certain details of the mechanism of their action and perform a comparative analysis
A. M. Magerramov; I. T. Nagieva
Green tea catechin induced phagocytosis can be blocked by catalase and an inhibitor of transient receptor potential melastatin 2 (TRPM2).
The major polyphenols in green tea, (-)-epigallocatechin and (-)-epigallocatechin gallate, have been shown to enhance the phagocytic activity of macrophage-like cells; however, the mechanism involved was not clarified. In this study, we have identified that the catechins induced phagocytosis can be blocked by catalase and an inhibitor of transient receptor potential melastatin 2. PMID:23896702
Monobe, Manami; Ema, Kaori; Tokuda, Yoshiko; Maeda-Yamamoto, Mari
A novel single-site manganese(II) complex of a pyridine derivative as a catalase mimetic for disproportionation of H2O2 in water.
A novel single site Mn(II) complex was successfully synthesized and tested in the aqueous disproportionation of hydrogen peroxide. The complex was found to be stable (HR-XAS) and exhibits catalase-like activity in neutral pH. Theoretical calculations suggested a reaction mechanism involving two complexes, changes in metal oxidation state and proton shuttling. PMID:23549197
Zienkiewicz, Ma?gorzata; Szlachetko, Jakub; Lothsch�tz, Christian; Hodorowicz, Maciej; Jab?o?ska-Wawrzycka, Agnieszka; S�, Jacinto; Barszcz, Barbara
6,6'-Bis(2-hydroxyphenyl)-2,2'-bipyridine manganese(III) complexes: a novel series of superoxide dismutase and catalase mimetics.
A series of novel manganese(III) complexes is described based on a 6,6'-bis(2-hydroxyphenyl)-2,2'-bipyridine template. These complexes show superoxide dismutase and catalase activity. The effect of the aromatic substitution pattern on the SAR is described. PMID:11378356
Giblin, G M; Box, P C; Campbell, I B; Hancock, A P; Roomans, S; Mills, G I; Molloy, C; Tranter, G E; Walker, A L; Doctrow, S R; Huffman, K; Malfroy, B
Localisation of Helicobacter pylori catalase in both the periplasm and cytoplasm, and its dependence on the twin-arginine target protein, KapA, for activity.
Helicobacter pylori induces a severe inflammatory response in the gastric mucosa. It is able to withstand the inflammatory response by producing proteins such as KatA and KapA. The C-terminus of KatA possesses a unique tetra-lysine motif not found in other catalases or other known protein sequences. Mutants deficient in this motif were constructed by site-directed mutagenesis. Cytoplasmic and periplasmic catalase activities were measured for the parental strain, a truncated KatA mutant (deficient in the unique C-terminal tetra-lysine motif) and a previously constructed KapA-deficient mutant (confirming previous observations regarding the possible periplasmic localisation of KatA). No differences were observed in the cytoplasmic catalase activities, however, the KapA-deficient mutant had approximately 5.5 times less catalase activity in the periplasmic extract when compared to the periplasmic preparations of either parental strain or KatA truncated mutant. N-terminal sequencing of KatA revealed no cleaved N-terminal signal peptide, indicating Sec-independent transport. These findings support previous reports that there is some form of interaction between KatA and KapA of H. pylori, an interaction which still needs to be characterised. PMID:14680712
Harris, Andrew G; Hazell, Stuart L
The Immunological Properties of Stroma-free Polyhemolysate Containing Catalase and Superoxide Dismutase Activities Prepared by Polymerized Bovine Stroma-free Hemolysate
Crosslinking of ultrapure hemoglobin, crystalline catalase, and superoxide dismutase resulted in a soluble nanodimensional complex of polyhemoglobin-catalase-superoxide dismutase. A less expensive and more convenient way is to crosslink bovine stroma-free hemolysate (stroma-free hemolysate) that already contains hemoglobin, catalase, and superoxide dismutase into polyhemoglobin with catalase and superoxide dismutase activities (stroma-free polyhemolysate) [21]. The objective of the present study is to evaluate the immunological properties of this stroma-free polyhemolysate. Each of three groups of rats received weekly subcutaneous injections of one of the stroma-free polyhemolysate, stroma-free hemolysate, and saline for four weeks. One week after the four cycles of weekly immunization, serum and plasma were collected for C3a complement activation tests and Ouchterlony antibody-antigen precipitation tests, respectively. Results show that stroma-free polyhemolysate retained significant antioxidant enzyme activity. The C3a complement activation test and Ouchterlony test show that four weekly subcutaneous injections of bovine stroma-free polyhemolysate did not result in any immunological reaction in rats when tested this way.
Zhu, Hongli; Du, Qianqian; Chen, Chao; Chang, Thomas Ming Swi
Upregulation of decorin by FXR in vascular smooth muscle cells
Decorin is a member of the family of small leucine-rich proteoglycans that are present in blood vessels and synthesized by vascular smooth muscle cells (VSMCs). Decorin plays complex roles in both normal vascular physiology and the pathogenesis of various types of vascular disorders. However, the mechanisms of regulation of decorin expression in vasculature are not clearly understood. Particularly little information is available about a role of nuclear receptors in the regulation of decorin expression. In the present study, we report that activation of vascular FXR by a specific ligand resulted in upregulation of decorin at the levels of both mRNA and protein. FXR appears to induce decorin expression at a transcriptional level because (1) upregulation of decorin mRNA expression was abolished by the treatment of a transcription inhibitor, actinomycin D; and (2) decorin promoter activity was significantly increased by activation of FXR. Functional analysis of human decorin promoter identified an imperfect inverted repeat DNA motif, IR8 (-2313TGGTCAtagtgtcaTGACCT-2294), as a likely FXR-responsive element that is involved in decorin regulation.
He Fengtian [Center for Pharmacogenetics, Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, 639 Salk Hall, Pittsburgh, PA 15261 (United States); Department of Biochemistry and Molecular Biology, Third Military Medical University, Chongqing 400038 (China); Zhang Qiuhong; Kuruba, Ramalinga; Gao Xiang; Li Jiang [Center for Pharmacogenetics, Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, 639 Salk Hall, Pittsburgh, PA 15261 (United States); Li Yong [Stem Cell Research Center, Children's Hospital of Pittsburgh, Pittsburgh, PA 15213 (United States); Gong Wei [Department of Biochemistry and Molecular Biology, Third Military Medical University, Chongqing 400038 (China); Jiang, Yu [Center for Pharmacogenetics, Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, 639 Salk Hall, Pittsburgh, PA 15261 (United States); Department of Biochemistry and Molecular Biology, Third Military Medical University, Chongqing 400038 (China); Xie Wen [Center for Pharmacogenetics, Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, 639 Salk Hall, Pittsburgh, PA 15261 (United States); Li Song [Center for Pharmacogenetics, Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, 639 Salk Hall, Pittsburgh, PA 15261 (United States)], E-mail: firstname.lastname@example.com
Aberrant upregulation of astroglial ceramide potentiates oligodendrocyte injury.
Oligodendroglial injury is a pathological hallmark of many human white matter diseases, including multiple sclerosis (MS) and periventricular leukomalacia (PVL). Critical regulatory mechanisms of oligodendroglia destruction, however, remain incompletely understood. Ceramide, a bioactive sphingolipid pivotal to sphingolipid metabolism pathways, regulates cell death in response to diverse stimuli and has been implicated in neurodegenerative disorders. We report here that ceramide accumulates in reactive astrocytes in active lesions of MS and PVL, as well as in animal models of demyelination. Serine palmitoyltransferase, the rate-limiting enzyme for ceramide de novo biosynthesis, was consistently upregulated in reactive astrocytes in the cuprizone mouse model of demyelination. Mass spectrometry confirmed the upregulation of specific ceramides during demyelination, and revealed a concomitant increase of sphingosine and a suppression of sphingosine-1-phosphate, a potent signaling molecule with key roles in cell survival and mitogenesis. Importantly, this altered sphingolipid metabolism during demyelination was restored upon active remyelination. In culture, ceramide acted synergistically with tumor necrosis factor, leading to apoptotic death of oligodendroglia in an astrocyte-dependent manner. Taken together, our findings implicate that disturbed sphingolipid pathways in reactive astrocytes may indirectly contri