In this study, we described an in-depth analytical characterization of the process of hydrolysis of polyester polymers catalyzed by a recently described cutinase (LCC WCCG ) and a homologue cutinase variant that was created and reported for the first time in this work (Tc_Cut2NV WCCG ). Model small molecules, aliphatic and aromatic polyesters as well as an aromatic-aliphatic co-polyester were investigated for their depolymerization with the chosen cutinases. RPLC-UV-MS analyses allowed us to monitor the water-soluble products released by hydrolysis. SEC-MS analysis allowed us to gain insight into the monomer composition of the species resulting from the enzymatic hydrolysis. We conclude that the mechanism of the enzymatic hydrolysis catalyzed by LCC WCCG and TC WCCG is similar and that in copolymer systems the enzymes selectively depolymerize the constituting monomers depending on their chemicophysical characteristics such as in the case illustrated here of aliphatic and aromatic monomers. The results and method proposed herein give insights into the mode of hydrolysis of these cutinases in polyester co-polymer systems, thereby providing information for the rational engineering of more selective and catalytically active enzymes. Furthermore, these results could be exploited as a starting point for developing analytical workflows in which the selectivity of enzymes is harnessed to characterize the copolymer distribution in a polymer sequence.