Development of a yeast whole-cell biocatalyst for MHET conversion into terephthalic acid and ethylene glycol
Abstract 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...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2022-12-01
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| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-022-02007-9 |
| _version_ | 1797973402360741888 |
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| author | Raphael Loll-Krippleber Victoria A. Sajtovich Michael W. Ferguson Brandon Ho Andrew R. Burns Brandon J. Payliss Joseph Bellissimo Sydney Peters Peter J. Roy Haley D. M. Wyatt Grant W. Brown |
| author_facet | Raphael Loll-Krippleber Victoria A. Sajtovich Michael W. Ferguson Brandon Ho Andrew R. Burns Brandon J. Payliss Joseph Bellissimo Sydney Peters Peter J. Roy Haley D. M. Wyatt Grant W. Brown |
| author_sort | Raphael Loll-Krippleber |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-04-11T04:03:42Z |
| format | Article |
| id | doaj.art-16c176dc105f499e87946f08f162cc78 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-04-11T04:03:42Z |
| publishDate | 2022-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-16c176dc105f499e87946f08f162cc782023-01-01T12:30:41ZengBMCMicrobial Cell Factories1475-28592022-12-0121111510.1186/s12934-022-02007-9Development of a yeast whole-cell biocatalyst for MHET conversion into terephthalic acid and ethylene glycolRaphael Loll-Krippleber0Victoria A. Sajtovich1Michael W. Ferguson2Brandon Ho3Andrew R. Burns4Brandon J. Payliss5Joseph Bellissimo6Sydney Peters7Peter J. Roy8Haley D. M. Wyatt9Grant W. Brown10Donnelly Centre for Cellular and Biomolecular Research, University of TorontoDonnelly Centre for Cellular and Biomolecular Research, University of TorontoDonnelly Centre for Cellular and Biomolecular Research, University of TorontoDonnelly Centre for Cellular and Biomolecular Research, University of TorontoDonnelly Centre for Cellular and Biomolecular Research, University of TorontoDepartment of Biochemistry, University of TorontoDonnelly Centre for Cellular and Biomolecular Research, University of TorontoDonnelly Centre for Cellular and Biomolecular Research, University of TorontoDonnelly Centre for Cellular and Biomolecular Research, University of TorontoDepartment of Biochemistry, University of TorontoDonnelly Centre for Cellular and Biomolecular Research, University of TorontoAbstract 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.https://doi.org/10.1186/s12934-022-02007-9PETMHETPlastic degradationMHETaseWhole-cell biocatalystSurface display |
| spellingShingle | Raphael Loll-Krippleber Victoria A. Sajtovich Michael W. Ferguson Brandon Ho Andrew R. Burns Brandon J. Payliss Joseph Bellissimo Sydney Peters Peter J. Roy Haley D. M. Wyatt Grant W. Brown Development of a yeast whole-cell biocatalyst for MHET conversion into terephthalic acid and ethylene glycol Microbial Cell Factories PET MHET Plastic degradation MHETase Whole-cell biocatalyst Surface display |
| title | Development of a yeast whole-cell biocatalyst for MHET conversion into terephthalic acid and ethylene glycol |
| title_full | Development of a yeast whole-cell biocatalyst for MHET conversion into terephthalic acid and ethylene glycol |
| title_fullStr | Development of a yeast whole-cell biocatalyst for MHET conversion into terephthalic acid and ethylene glycol |
| title_full_unstemmed | Development of a yeast whole-cell biocatalyst for MHET conversion into terephthalic acid and ethylene glycol |
| title_short | Development of a yeast whole-cell biocatalyst for MHET conversion into terephthalic acid and ethylene glycol |
| title_sort | development of a yeast whole cell biocatalyst for mhet conversion into terephthalic acid and ethylene glycol |
| topic | PET MHET Plastic degradation MHETase Whole-cell biocatalyst Surface display |
| url | https://doi.org/10.1186/s12934-022-02007-9 |
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