Discovery and mechanism-guided engineering of BHET hydrolases for improved PET recycling and upcycling.

ABSTRACT: Although considerable research achievements have been made to address the plastic crisis using enzymes, their applications are limited due to incomplete degradation and low efficiency. Herein, we report the identification and subsequent engineering of BHETases, which have the potential to improve the efficiency of PET recycling and upcycling. Two BHETases (ChryBHETase and BsEst) are identified from the environment via enzyme mining. Subsequently, mechanism-guided barrier engineering is employed to yield two robust and thermostable ΔBHETases with up to 3.5-fold enhanced k_cat/K_M than wild-type, followed by atomic resolution understanding. Coupling ΔBHETase into a two-enzyme system overcomes the challenge of heterogeneous product formation and results in up to 7.0-fold i

SUBMITTER: Li A 

PROVIDER: S-EPMC10344914 | biostudies-literature | 2023 Jul

REPOSITORIES: biostudies-literature