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

Anni Li, Yijie Sheng, Haiyang Cui, Minghui Wang, Luxuan Wu, Yibo Song, Rongrong Yang, Xiujuan Li () and He Huang ()
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Anni Li: Nanjing Normal University
Yijie Sheng: Nanjing Normal University
Haiyang Cui: RWTH Aachen University
Minghui Wang: Nanjing Normal University
Luxuan Wu: Nanjing Normal University
Yibo Song: Nanjing Normal University
Rongrong Yang: Nanjing Normal University
Xiujuan Li: Nanjing Normal University
He Huang: Nanjing Normal University

Nature Communications, 2023, vol. 14, issue 1, 1-16

Abstract: 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 kcat/KM 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 improved TPA production than seven state-of-the-art PET hydrolases, under the conditions used here. Finally, we employ a ΔBHETase-joined tandem chemical-enzymatic approach to valorize 21 commercial post-consumed plastics into virgin PET and an example chemical (p-phthaloyl chloride) for achieving the closed-loop PET recycling and open-loop PET upcycling.

Date: 2023
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DOI: 10.1038/s41467-023-39929-w

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