In:
Angewandte Chemie International Edition, Wiley, Vol. 62, No. 14 ( 2023-03-27)
Abstract:
PETase displays great potential in PET depolymerization. Directed evolution has been limited to engineer PETase due to the lack of high‐throughput screening assay. In this study, a novel fluorescence‐based high‐throughput screening assay employing a newly designed substrate, bis (2‐hydroxyethyl) 2‐hydroxyterephthalate (termed BHET‐OH), was developed for PET hydrolases. The best variant DepoPETase produced 1407‐fold more products towards amorphous PET film at 50 °C and showed a 23.3 °C higher T m value than the PETase WT. DepoPETase enabled complete depolymerization of seven untreated PET wastes and 19.1 g PET waste (0.4 % W enzyme /W PET ) in liter‐scale reactor, suggesting that it is a potential candidate for industrial PET depolymerization processes. The molecular dynamic simulations revealed that the distal substitutions stabilized the loops around the active sites and transmitted the stabilization effect to the active sites through enhancing inter‐loop interactions network.
Type of Medium:
Online Resource
ISSN:
1433-7851
,
1521-3773
DOI:
10.1002/anie.202218390
Language:
English
Publisher:
Wiley
Publication Date:
2023
detail.hit.zdb_id:
2011836-3
detail.hit.zdb_id:
123227-7
