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Fluorescence-activated Screening of Polyester-depolymerizing Enzymes Based on Pseudo-PET Polythioester Plastics
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  • Xiaoqiang Chen,
  • Yuanbo Wang,
  • Jie Zhou,
  • Canhao Qiu,
  • Lei Wang,
  • Shiyue Zheng,
  • Junqian Peng,
  • Sheng Lu,
  • Fang Wang,
  • Ziyi Yu,
  • Weiliang Dong,
  • Min Jiang
Xiaoqiang Chen
Nanjing Tech University State Key Laboratory of Materials-Oriented Chemical Engineering

Corresponding Author:[email protected]

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Yuanbo Wang
Nanjing Tech University State Key Laboratory of Materials-Oriented Chemical Engineering
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Jie Zhou
Nanjing Tech University College of Biotechnology and Pharmaceutical Engineering
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Canhao Qiu
Nanjing Tech University College of Biotechnology and Pharmaceutical Engineering
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Lei Wang
Nanjing Tech University State Key Laboratory of Materials-Oriented Chemical Engineering
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Shiyue Zheng
Nanjing Tech University State Key Laboratory of Materials-Oriented Chemical Engineering
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Junqian Peng
Nanjing Tech University State Key Laboratory of Materials-Oriented Chemical Engineering
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Sheng Lu
Nanjing Tech University State Key Laboratory of Materials-Oriented Chemical Engineering
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Fang Wang
Nanjing Tech University State Key Laboratory of Materials-Oriented Chemical Engineering
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Ziyi Yu
Nanjing Tech University State Key Laboratory of Materials-Oriented Chemical Engineering
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Weiliang Dong
Nanjing Tech University College of Biotechnology and Pharmaceutical Engineering
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Min Jiang
Nanjing Tech University College of Biotechnology and Pharmaceutical Engineering
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Abstract

Fluorescence-based high-throughput screening approaches facilitate the discovery of enzymes and microorganisms for polyethylene terephthalate (PET) depolymerization and recycling. However, the traditional method of activating the fluorescence signal by cleaving the ester bond on small molecule probes has limited ability in detecting enzymatic activity towards polymeric substrates. This study proposes a novel fluorescence-based screening strategy that detects the release of sulfhydryl groups during the depolymerization of pseudo-PET polythioesters by polyesterases. The strategy successfully identifies the polyester-depolymerizing activity of leaf-branch compost cutinase (LCC ICCG), while porcine liver esterase (PLE) only hydrolyses small molecular substrates. When combined with a droplet microfluidic system, the strategy enables high-throughput screening of LCC ICCG. The study also demonstrates that screening for polyester-depolymerizing bacteria can be performed via a microplate reader platform. The new screening approach offers an efficient method for identifying enzymes and microbial resources for depolymerizing polyester-like plastics.
Submitted to AIChE Journal
31 Jan 2024Review(s) Completed, Editorial Evaluation Pending
01 Feb 2024Reviewer(s) Assigned
22 Apr 2024Submission Checks Completed
22 Apr 2024Assigned to Editor
22 Apr 2024Reviewer(s) Assigned
29 May 2024Editorial Decision: Revise Major
10 Jun 2024Submission Checks Completed
10 Jun 2024Assigned to Editor
10 Jun 2024Review(s) Completed, Editorial Evaluation Pending
06 Jul 2024Editorial Decision: Accept