Above the Tg, the amorphous regions of polymers exhibit increased molecular mobility and are more susceptible to enzymatic attacks due to the greater free volume available for diffusion, thereby facilitating depolymerization. Moreover, different strategies to increase the surface area were applied and are described in the SI Figures and. Two methods were used to characterize the hydrolysis products: RPLC-UV-MS to monitor the release of dicarboxylic acid monomers in the aqueous reaction mixture (3 for PES3 and 2 for PES1) and SEC-UV to analyze the remaining polymer material after centrifugation, sample evaporation, and re-dissolution in HFIP. Details of the RPLC and SEC analysis are presented in Figures S14, S15 and S17. Figure illustrates the molecular weight distribution (relative to polystyrene standards) as Log MW plots obtained for the depolymerization reactions of PES1 by LCC WCCG and Tc_Cut2NV WCCG after different reaction times. Negative control experiments (i.e., devoid of enzyme, black trace) were also performed. We observed complete depolymerization of PES1 with LCC WCCG after 30 min (yellow trace) as the polymer distribution area disappeared. In the case of Tc_Cut2NV WCCG (blue trace), the PES1 polymer peak decreased in 30 min and the complete depolymerization was observed within less than 24 h. The process went hand in hand with the progressive formation of 2 as observed by RPLC-MS and the visual observation of the complete degradation of PES1 in the reaction mixture.