Dataset Information

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 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.

SUBMITTER: Loll-Krippleber R

PROVIDER: S-EPMC9805092 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature

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Publications

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

Loll-Krippleber Raphael R Sajtovich Victoria A VA Ferguson Michael W MW Ho Brandon B Burns Andrew R AR Payliss Brandon J BJ Bellissimo Joseph J Peters Sydney S Roy Peter J PJ Wyatt Haley D M HDM Brown Grant W GW

Microbial cell factories 20221231 1

<h4>Background</h4>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 ...[more]

PMID: 36587193

Similar Datasets

Project description:Komagataeibacter xylinus strains DSM 2004 and DSM 46604 were evaluated for their ability to grow and produce bacterial cellulose (BC) upon cultivation on terephthalic acid (TA) and ethylene glycol (EG), which are monomers of the petrochemical-derived plastic polyethylene terephthalate (PET). Both strains were able to utilize TA, EG, and their mixtures for BC synthesis, with different performances. K. xylinus DSM 2004 achieved higher BC production from TA (0.81 ± 0.01 g/L), EG (0.64 ± 0.02 g/L), and TA + EG mixtures (0.6 ± 0.1 g/L) than strain DSM 46604. The latter was unable to utilize EG as the sole carbon source and reached a BC production of 0.16 ± 0.01 g/L and 0.23 ± 0.1 g/L from TA alone or TA + EG mixtures, respectively. Further supplementing the media with glucose enhanced BC production by both strains. During cultivation on media containing TA and EG, rapid pH drop due to metabolization of EG into acidic compounds led to some precipitation of TA that was impregnated into the BC pellicles. An adaptation of the downstream procedure involving BC dissolution in NaOH was used for the recovery of pure BC. The different medium composition tested, as well as the downstream procedure, impacted the BC pellicles' physical properties. Although no variation in terms of the chemical structure were observed, differences in crystallinity degree and microstructure of the produced BC were observed. The BC produced by K. xylinus DSM 2004 had a higher crystallinity (19-64%) than that of the strain DSM 46604 (17-53%). Moreover, the scanning electron microscopy analysis showed a higher fiber diameter for K. xylinus DSM 2004 BC (46-56 nm) than for K. xylinus DSM 46604 (37-49 nm). Dissolution of BC in NaOH did not influence the chemical structure; however, it led to BC conversion from type I to type II, as well as a decrease in crystallinity. These results demonstrate that PET monomers, TA and EG, can be upcycled into a value-added product, BC, presenting an approach that will contribute to lessening the environmental burden caused by plastic disposal in the environment.

Dataset Information

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

Background

Results

Conclusions

Publications

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

Similar Datasets

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