The molecular docking poses in the productive binding modes show the influence of the catalytic histidine residue (H209 for Tc_Cut2NV WCCG and H242 for LCC WCCG ) in the protonation of the oxygen atom of the carbonyl involved in the hydrolysis reaction. Notably, the introduced tryptophan mutation (F210W for Tc_Cut2NV WCCG or F243W for LCC WCCG ) must influence the positioning of the low molar mass substrates in the active site through the possible formation of a hydrogen bond between the NH moiety of the tryptophan residue and the oxygen atom of the second carbonyl (i.e., not involved in the hydrolysis) of the substrate (See Fig. ). As predicted by the docking studies, both Tc_Cut2NV WCCG or LCC WCCG could hydrolyze the test low molar mass substrates. After 1 or 24 h, we detected the formation by LC-MS of monomer 3 or 5 as the products of the depolymerization of model substrates 8 or 9, respectively. Within the investigated timeframe (up to 24 h) for the depolymerization of 8 or 9, LCC WCCG produced a greater amount of the monomer 3 or 5 (near 100% hydrolysis) compared with Tc_Cut2NV WCCG (SI, Fig. ; around 80% of hydrolysis). Notably, using Tc_Cut2NV WCCG , we detected the formation of partially hydrolyzed intermediates, thus maintaining one of the two ester moieties (compound 8a and 9a of SI Fig. ). In contrast, both intermediates were consumed by LCC WCCG after 30 min of reaction. These observations indicate a different reactivity between Tc_Cut2NV WCCG and LCC WCCG , with the latter enzyme being more catalytically active. Moreover, the hydrolysis of the aliphatic low-molar-mass substrate 9 catalyzed by LCC WCCG was significantly faster than that of the aromatic low-molar-mass substrate 8 within the first 100 min of the reaction.