Development of a yeast whole-cell biocatalyst for MHET conversion into terephthalic acid and ethylene glycol

Microbial Cell Factories

Research

Brandon J. Payliss¹  Haley D. M. Wyatt¹  Sydney Peters²  Grant W. Brown²  Michael W. Ferguson²  Brandon Ho²  Raphael Loll-Krippleber²  Joseph Bellissimo²  Andrew R. Burns³  Peter J. Roy⁴  Victoria A. Sajtovich⁵ 

[1] Department of Biochemistry, University of Toronto, Toronto, Canada;Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Canada;Department of Biochemistry, University of Toronto, Toronto, Canada;Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Canada;Department of Molecular Genetics, University of Toronto, Toronto, Canada;Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Canada;Department of Molecular Genetics, University of Toronto, Toronto, Canada;Department of Pharmacology and Toxicology, University of Toronto, Toronto, Canada;Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Canada;Max Planck Institute for Terrestrial Microbiology, Marburg, Germany;

关键词: PET;    MHET;    Plastic degradation;    MHETase;    Whole-cell biocatalyst;    Surface display;    Yeast;    Saccharomyces cerevisiae;   

DOI  :  10.1186/s12934-022-02007-9

 received in 2022-11-01, accepted in 2022-12-20,  发布年份 2022

来源: Springer

【 摘 要 】

BackgroundOver the 70 years since the introduction of plastic into everyday items, plastic waste has become an increasing problem. With over 360 million tonnes of plastics produced every year, solutions for plastic recycling and plastic waste reduction are sorely needed. Recently, multiple enzymes capable of degrading PET (polyethylene terephthalate) plastic have been identified and engineered. In particular, the enzymes PETase and MHETase from Ideonella sakaiensis depolymerize PET into the two building blocks used for its synthesis, ethylene glycol (EG) and terephthalic acid (TPA). Importantly, EG and TPA can be re-used for PET synthesis allowing complete and sustainable PET recycling.ResultsIn this study we used Saccharomyces cerevisiae, a species utilized widely in bioindustrial fermentation processes, as a platform to develop a whole-cell catalyst expressing the MHETase enzyme, which converts monohydroxyethyl terephthalate (MHET) into TPA and EG. We assessed six expression architectures and identified those resulting in efficient MHETase expression on the yeast cell surface. We show that the MHETase whole-cell catalyst has activity comparable to recombinant MHETase purified from Escherichia coli. Finally, we demonstrate that surface displayed MHETase is active across a range of pHs, temperatures, and for at least 12 days at room temperature.ConclusionsWe demonstrate the feasibility of using S. cerevisiae as a platform for the expression and surface display of PET degrading enzymes and predict that the whole-cell catalyst will be a viable alternative to protein purification-based approaches for plastic degradation.

【 授权许可】

CC BY   
© The Author(s) 2022

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