Hydrophobic cell surface display system of PETase as a sustainable biocatalyst for PET degradation

Jia, Yantao; Samak, Nadia A.; Hao, Xuemi; Chen, Zheng; Wen, Qifeng; Xing, Jianmin

doi:10.3389/fmicb.2022.1005480

Cited by 14 publications

(12 citation statements)

References 35 publications

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“…Surface display systems for PETase show little loss of enzyme activity over 7 days [ 28 , 44 ], whereas soluble PETase loses activity more rapidly. We compared the activity of surface displayed MHETase to soluble purified MHETase after incubating for up to 12 days at room temperature in phosphate buffer.…”

Section: Resultsmentioning

confidence: 99%

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

et al. 2022

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Background Over 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. Results In 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. Conclusions We 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.

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Section: Resultsmentioning

confidence: 99%

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

et al. 2022

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“…The PET degradation efficiency of the co-display systems was significantly increased compared to E. coli cells that solely displayed Fast-PETase on the surfaces, proving the feasibility and utility of co-display approach with E. coli system. Because of the use of different PET materials and PETase enzymes for degradation tests (Chen et al 2022 ; Gercke et al 2021 ; Jia et al 2022 ; Zhu et al 2022 ), it was impossible for us to make a direct comparison with other reported systems. Nevertheless, over 15% degradation of amorphous PET in 24 h was the highest value to date.…”

Section: Discussionmentioning

confidence: 99%

“…1 A). To overcome the limitations of using free enzymes, such as instability and time-consuming purification process, various whole cell systems have been constructed for PET biodegradation using the PETases from different sources (Chen et al 2022 , 2020 ; Gercke et al 2021 ; Jia et al 2022 ; Zhu et al 2022 ). However, the efficiency of PET biodegradation by these systems is still lower than realistic applications.…”

Section: Introductionmentioning

confidence: 99%

Constructing Escherichia coli co-display systems for biodegradation of polyethylene terephthalate

Hu,

Chen

2023

Bioresour. Bioprocess.

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Background The accumulation of fast-growing polyethylene terephthalate (PET) wastes has posed numerous threats to the environments and human health. Enzymatic degradation of PET is a promising approach for PET waste treatment. Currently, the efficiency of various PET biodegradation systems requires further improvements. Results In this work, we engineered whole cell systems with co-display of strong adhesive proteins and the most active PETase for PET biodegradation in E. coli cells. Adhesive proteins of cp52k and mfp-3 and Fast-PETase were simultaneously displayed on the surfaces of E. coli cells, and the resulting cells displaying mfp-3 showed 50% increase of adhesion ability compared to those without adhesive proteins. Consequently, the degradation rate of E. coli cells co-displaying mfp-3 and Fast-PETase for amorphous PET exceeded 15% within 24 h, exhibiting fast and thorough PET degradation. Conclusions Through the engineering of co-display systems in E. coli cells, PET degradation efficiency was significantly increased compared to E. coli cells with sole display of Fast-PETase and free enzyme. This feasible E. coli co-display system could be served as a convenient tool for extending the treatment options for PET biodegradation. Graphical Abstract

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“…Surface display systems for PETase show little loss of enzyme activity over 7 days [25,40], whereas soluble PETase loses activity more rapidly. We compared the activity of surface displayed MHETase to soluble purified MHETase after 12 days at room temperature in phosphate buffer.…”

Section: The Mhetase Whole-cell Catalyst Is Stable To Alkaline Ph Tem...mentioning

confidence: 99%

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

Loll-Krippleber

Sajtovich

Ferguson

et al. 2022

Preprint

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Background. Over 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 teraphthalate) 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. Results. In this study, we used Saccharomyces cerevisiae as a platform to develop a whole-cell catalyst expressing the MHETase enzyme, which converts MHET (monohydroxyethyl terephthalate) 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 stable to pH, temperature, and for at least 12 days at room temperature. Conclusions. We 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 viable alternatives to protein purification-based approaches for plastic degradation.

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Hydrophobic cell surface display system of PETase as a sustainable biocatalyst for PET degradation

Cited by 14 publications

References 35 publications

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

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

Constructing Escherichia coli co-display systems for biodegradation of polyethylene terephthalate

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

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