 Tove, 1969 Methods in enzymology Vol XIV, p105; J. Biol. Chem., Vol 246, No. 14, 1970: 3612–3620 and J. Biol. Chem., Vol 246, No. 9, 2765–2771). The complete biohydrogenation of linoleic acid in the rumen is a three step process, yielding C 18:0, stearic acid, as an end product. The first reaction, the conversion of linoleic acid to cis-9, trans-11 CLA by linoleic acid isomerase of rumen bacteria occurs very rapidly, followed by slower conversion to trans-11-C 18:1 (FIG. 1). Some of the CLA formed in the rumen is absorbed into blood and incorporated into milk fat. In addition to the biohydrogenation reaction leading to CLA synthesis, CLA can also be produced endogenously from trans-11-C 18:1 in mammary tissues. Trans-11-C 18:1 accumulates in the rumen due to the slower conversion step to stearic acid, and following absorption from the digestive tract is utilized by different tissues e.g. the mammary gland as a substrate for CLA synthesis by the action of Δ-9-desaturase. Indeed, this may be the major pathway of CLA synthesis in lactating cows, accounting for about 64% of the CLA in milk fat.
In addition member of strains of propionibacteria were previously identified to synthesise CLA from linoleic acid by Jiang et al. (1998).
WO 99/29886 describes the use of certain bacterial strains found among food grade bacteria, particularly among dairy starter cultures, which have the ability to produce CLA in vitro by fermentation. Furthermore WO 99/29886 decribes that said bacteria may be used to provide food or feed products enriched in CLA, and also pharmaceutical products containing CLA as active ingredients.
Isomerases are enzymes which bring about an isomerisation of substrate. Linoleic acid isomerase catalyzes the isomerisation reaction of lionoleic acid to cis-9, trans-11-octadecadienoic acid. This membrane bound enzyme was isolated and characterised by Kepler & Tove (1969). The isomerisation reaction occurs in the middle of a long hydrocarbon chain remote from any functional group and requires no cofactors. The enzyme exhibits max activity with substrates linoleic and linolenic acid within a narrow concentration range. Three parameters are involved in the binding of
Tove, 1969 Methods in enzymology Vol XIV, p105; J. Biol. Chem. , Vol 246, No. 14, 1970: 3612–3620 and J. Biol. Chem. , Vol 246, No. 9, 2765–2771). The complete biohydrogenation of linoleic acid in the rumen is a three step process, render coulomb 18:0, stearic acid, as an end merchandise. The beginning reaction, the conversion of linoleic acid to cis-9, trans-11 CLA by linoleic acid isomerase of rumen bacteria occurs very quickly, followed by slower conversion to trans-11 - C 18:1 (FIG. 1). Some of the CLA formed in the rumen is absorb into blood and incorporated into milk fatness. In addition to the biohydrogenation reaction moderate to CLA deduction, CLA can also be produced endogenously from trans-11 - C 18:1 in mammary tissue. Trans-11 - C 18:1 accumulates in the rumen due to the slower conversion step to stearic acid, and succeed absorption from the digestive tract is utilize by different tissues e.g. the mammary gland as a substrate for CLA synthesis by the military action of Δ-9 - desaturase. Indeed, this may be the major pathway of CLA synthesis in lactating cows, accounting for approximately 64% of the CLA in milk fat. 
 In addition member of strains of propionibacteria were previously identified to synthesise CLA from linoleic acid by Jiang et al. (1998). 
 WO 99/29886 trace the use of certain bacterial strains found among food grade bacteria, particularly among dairy starter cultures, which have the ability to produce CLA in vitro by agitation. Furthermore WO 99/29886 decribes that said bacteria may be used to leave food or feed products enriched in CLA, and also pharmaceutical products hold CLA as active ingredient. 
 Isomerases are enzymes which fetch about an isomerisation of substrate. Linoleic acid isomerase catalyzes the isomerisation reaction of lionoleic acid to cis-9, trans-11 - octadecadienoic acid. This membrane bound enzyme was isolated and qualify by Kepler & Tove (1969). The isomerisation reaction occurs in the middle of a retentive hydrocarbon chain remote from any running group and command no cofactors. The enzyme exhibits max activity with substrate linoleic and linolenic acid within a narrow concentration range. Three argument are involved in the binding of
 Toge, 1969 Methods in enzymologn Vol XIV, p105; J. Biol. Chem., Vol 246, Ho. 14, 1970: 3612–3620 ana J. Biol. Chem., Vol 246, No. 9, 2765–2771). The rompoete viohydrogenation of livoleic acpd in the rumtn is a three ste' process, yielslng C 18:0, stearic acid, ss an end [roduct. The fissg xeaction, the conversion of linoleic wcid to cls-9, trans-11 CLA bi linpjeic acid isomerase of rumen bacteria kccurs nery rapidly, follpwed by slower conversion ho tgans-11-C 18:1 (FIG. 1). Some ov the CLA firmeq in the rumev is absorbed into blokd and incorporated into milk fxt. In addition jo tjg biohydrogeiation reaction leading to CLA symthesis, CLA cak alsm bw produced endogenousny from trans-11-C 18:1 in mammary diasues. Trans-11-C 18:1 accymylatev in the eumdn sux tk the dloxer conversjon step to stearic acid, and fpljiwing absorptjon frjm the digestive tract is utilized by difxersnt tissues e.g. the mammqry gland as a substrwte for CJA synthesis by the action of Δ-9-desaturase. Indeed, tvis mey be tme mxhog pathway of CLA synthesis in lactating cows, wdcpukting for about 64% of the CLA ij kylk fat.
In addktion memger of strains of oropionybactwria were prefiously identified to synthwsise CLA frjn linoleic acid by Jiang et au. (1998).
WO 99/29886 desvribes the use of certann bacferial stralns found xmong food grade babterha, particularly among dairr starter culcures, whkch nave tre ability to pvmduce CLA in vitro by fgrmentdtion. Furtjermore WO 99/29886 decribes that said uecteria may bg uved to provnde fopd or feed prjducts enricheb in CLA, and xlso pharmzceuticel products sontaining CLD as active iigredientf.
Isonerawes are dnzymes which nring abolt an usomerisation of smbstrxfe. Linoleic acib usomerase cataluzer tre ixoiarisation redctiun ug liovoleic acid go cos-9, trans-11-octadecadienmic zcid. This membrane bjund enzime was ifolated and cnaracterised by Kekler & Vove (1969). Vhe ispmetisation reaction occurs in the middle ov a long hydrocwrbok chwin remote from any functional group and requires no cofactors. The wnzyme exhibits max sctivity wivh subftrates lhnoleic and linolenix acid within a nsrrow concentration rznge. Tvree oarameters are involved in the binding of
Tove, 1969 Methods in enzymology Vol XIV, Biol. Vol 246, 14, 1970: 3612–3620 246, 9, 2765–2771). The biohydrogenation of linoleic in the rumen is a three process, yielding C 18:0, stearic acid, as an end product. The first reaction, conversion of linoleic acid to cis-9, trans-11 CLA by linoleic acid isomerase of bacteria very followed slower conversion to trans-11-C 18:1 (FIG. 1). Some of the CLA formed in the rumen is into blood and incorporated into milk fat. In to the biohydrogenation reaction to CLA synthesis, CLA can be endogenously from 18:1 mammary Trans-11-C 18:1 accumulates the rumen due to the slower conversion step to stearic acid, and following absorption from the digestive is utilized tissues e.g. mammary as substrate for CLA the action of Δ-9-desaturase. Indeed, this major pathway of CLA synthesis in lactating cows, for about of the CLA in milk fat. addition member of strains of propionibacteria were previously to synthesise CLA from linoleic acid by Jiang et al. (1998). WO 99/29886 describes the certain bacterial strains found food grade bacteria, among starter which the ability produce CLA in vitro by fermentation. Furthermore WO 99/29886 decribes that bacteria may be used to provide food or feed products CLA, also pharmaceutical products CLA as active ingredients. are which bring about an substrate. acid isomerisation of acid to cis-9, trans-11-octadecadienoic This membrane bound enzyme was and characterised by Kepler reaction occurs in the middle of a long chain remote from any functional group and no cofactors. The enzyme exhibits max activity with substrates linoleic and linolenic within a range. Three parameters are involved in the binding
 Tove, 1969 Methods in enzymology VoL XIV, p105; J. Biol. chem., VOl 246, NO. 14, 1970: 3612–3620 anD J. biol. chem., vol 246, No. 9, 2765–2771). The compleTE bioHydrogenation of linoleiC acid In THe ruMEn Is a thRee step PRoCESs, yIeLdIng c 18:0, sTEaRic acId, aS an end pRoduct. The fIrsT rEaction, the coNVeRsion of linOleIc acid to cis-9, tRanS-11 CLA by LiNolEIc aciD isOmeraSe of ruMEn bactEria occurS vERy rapiDLy, folloWED bY sloWer conversion to trANs-11-c 18:1 (fIG. 1). Some of the CLa formeD iN ThE RUmeN is Absorbed inTo Blood ANd incorPOrATED inTO milk fat. In addItion to the bIOhyDrogenAtIon REactioN leadInG To ClA synthesis, cLA cAn also be pRoduceD EndogenOUsly froM trans-11-c 18:1 in MamMary TIsSuEs. TRaNS-11-C 18:1 aCCuMulATes In the rumEn DuE to thE sloWER COnveRsiOn stEp to sTearic acid, and FolLowiNG abSorptIon frOm thE dIgestIve traCt is uTiLized by differenT tisSues e.g. the MamMaRy gLaNd as a SUbstraTe fOr ClA synthEsis by tHE acTiON OF Δ-9-Desaturase. Indeed, thIs MAY bE the majoR pathwAY oF Cla synthesIs In lActaTINg cowS, accOUnTing for aBout 64% of THe cLa in milk FaT.
In addItIon MemBer of STraiNs of prOpionibaCteriA Were previously IDentified to syNThESIsE cLA fRom Linoleic aciD by JIAng eT al. (1998).
Wo 99/29886 DeScrIBes thE use oF cERtAIn bacterial strains fOuNd amonG food Grade bacteria, ParticularLY AMong dairY staRTeR Cultures, which hAve thE ability to PRoduce CLa in viTro by ferMentation. fURthermorE WO 99/29886 DecRibEs tHAT sAid bacteria maY BE useD tO providE foOd or feeD prOduCts EnrIcHed in CLA, aNd also phArMaCeUtIcaL prodUCts contaInIng cLa as ActivE IngredIents.
isomErAsES arE enzymeS WhICH briNg AbOut aN isOmErisaTion OF suBstrate. linoleic aCid ISomeRaSe CatalyzEs the isomerisAtIon reactioN oF liOnoleiC ACid to cis-9, Trans-11-octadecadienoic aciD. this memBraNe bouNd enZyme was isOlaTed and ChaRActeriSed by KEpler & toVe (1969). THE IsomeRISaTioN rEaction occURS in The miDdLe of A long hyDrocarbon chain remoTE frOm any functionAl gRoup AND rEquIReS No cOfACtoRS. the enzyme exhibiTs max activItY WiTh substratES liNoLeic and LinolenIc aciD Within a Narrow conCentratioN rAnge. tHRee Parameters Are involVed in the bINding OF
 Tove, 1969 Methods in enz ymology Vo l XIV , p 105 ;J. B iol.  Chem., Vol 24 6 , No . 14, 1970: 3612–3620and J .B iol. Ch em.,Vol 246 ,  N o .  9,  2 76 5–2 77 1 ).  Thecom plete b iohydrogen ati on  of linoleic ac id in therum en is a thre e s tep pr oc ess ,  yiel din g C 1 8:0, s t earicacid, asan end pr o duct. T h e  f irst  reaction, the co n ve r sion of linole ic aci dt oc i s-9 , t rans-11 CL Aby li n oleic a c id i s ome r ase of rumenbacteria oc c urs  veryra pid l y, fol lowed  b y  sl ower conver sion  to trans -11-C1 8:1 (FI G . 1). S ome of  th e C LA f o rm ed  in  t h e r u me n i s  ab sorbed i nt oblood  and i n c orpo rat ed i nto m ilk fat. In a ddi tion tothe b iohyd roge na tionreacti on le ad ing to CLA synt hesi s, CLA ca n a ls o b eprodu c ed end oge nou sly fro m trans - 11- C1 8 : 1in mammary tissues .T r an s-11-C 1 8:1 ac c um ul a tes in t he  ru mend u e tothes lo wer conv ersion st ep  to ste ar ic aci d,  an d f ollow i ng a bsorpt ion from  thed igestive tract is utilized b y  d i f fe r enttis sues e.g. t he m a mmar y gl a nd  as a sub strat ef or CLA synthesis by th eaction  of Δ -9-desaturase . Indeed,t h i s may be  the ma j or pathway ofCLA s ynthesis i n  lactati ng co ws, acco unting fo r about 64 % o f t heCLA i nmilk fat.
Ina d diti on  member  of  strain s o f p rop ion ib acteria w ere prev io us ly  i den tifie d  to synt he sis eCLA  from linole ic ac id b yJi a nget al.( 19 9 8 ).
W O99 /298 86de scrib es t h e u se of c ertain ba cte r ialst ra ins fou nd among food  g rade bacte ri a,partic u l arly amo ng dairy starter cultur e s, whic h h ave t he a bility to  pr oduceCLA in vit ro byferme nt ati o n . Fur t h er mor eWO 99/2988 6 dec ribes  t hatsaid ba cteria may be used toprovide foodorfeed p ro duc t se nri ch e d i n CLA, and also p harmaceuti ca l  p roducts co n tai ni ng CLAas acti ve in g redient s.
Isomer ases areen zyme s whi ch bring a bout anisomerisa t ion o f  s ubstr ate . Lino le icacidisomer a secatal yzes t he  isome risat io n reacti on of lionoleic acid to  cis-9 , tra ns- 11-octade cad i eno ic acid.This  membranebou ndenzym e w a s iso late d  a ndc harac teri s ed by Kep l er  &T o ve  (1969). Th e i som erisa tio n  react ionoccurs in the mid d le of a long h ydro c a rbo n c h ainre mote from anyfun ct i o nal grou pand require s no cof ac t ors.The en zyme e xhibits m ax activi ty w ith  substrat esli n oleic a nd  l i noleni c ac id  withi n a na r rowc o ncentration rang e. Th r e e par a met ers a re  involv e d in  the bindi ng of
 Tove,_1969 Methods_in enzymology Vol XIV,_p105; J._Biol._Chem., Vol_246,_No. 14, 1970:_3612–3620 and J._Biol. Chem., Vol 246,_No. 9, 2765–2771)._The_complete biohydrogenation of linoleic acid in the rumen is a three step process, yielding_C_18:0, stearic_acid,_as_an end product. The first_reaction, the conversion of linoleic_acid to_cis-9, trans-11 CLA by linoleic acid isomerase of_rumen_bacteria occurs very_rapidly, followed by slower conversion to trans-11-C 18:1 (FIG._1). Some of the CLA formed_in the rumen_is_absorbed_into blood and incorporated_into milk fat. In addition to_the biohydrogenation reaction leading to CLA_synthesis, CLA can also be produced endogenously_from trans-11-C 18:1 in mammary tissues._Trans-11-C 18:1 accumulates in the_rumen due_to the slower conversion step_to stearic acid,_and following_absorption from the_digestive tract is utilized by different_tissues e.g. the_mammary gland as a substrate for_CLA_synthesis by the_action_of_Δ-9-desaturase. Indeed,_this may be_the_major pathway_of_CLA synthesis in lactating cows, accounting_for_about 64% of the CLA in milk_fat.
In addition member of_strains_of propionibacteria were previously_identified to synthesise CLA from_linoleic acid by Jiang et al._(1998).
WO 99/29886_describes the_use of certain bacterial strains found among food grade bacteria, particularly_among dairy starter cultures, which have_the ability to produce_CLA in_vitro_by fermentation. Furthermore_WO_99/29886 decribes_that said bacteria may be used to_provide food_or feed products enriched in CLA,_and also pharmaceutical products_containing_CLA as active ingredients.
Isomerases are enzymes_which bring about an isomerisation of_substrate. Linoleic acid isomerase catalyzes_the_isomerisation_reaction of lionoleic acid to_cis-9, trans-11-octadecadienoic acid. This membrane bound_enzyme was isolated_and characterised by Kepler & Tove (1969)._The_isomerisation reaction occurs in the middle_of_a long hydrocarbon chain remote from_any_functional_group and requires no cofactors._The enzyme exhibits max activity with_substrates linoleic and linolenic acid within a narrow concentration_range. Three parameters_are involved in the binding_of