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Surface display as a functional screening platform for detecting enzymes active on PET
Poly(ethylene terephthalate) (PET) is the world’s most abundant polyester plastic, and its ongoing accumulation in nature is causing a global environmental problem. Currently, the main recycling processes utilize thermomechanical or chemical means, resulting in the deterioration of the mechanical pr...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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BioMed Central
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8088578/ https://www.ncbi.nlm.nih.gov/pubmed/33933097 http://dx.doi.org/10.1186/s12934-021-01582-7 |
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author | Heyde, Sophia A. H. Arnling Bååth, Jenny Westh, Peter Nørholm, Morten H. H. Jensen, Kenneth |
author_facet | Heyde, Sophia A. H. Arnling Bååth, Jenny Westh, Peter Nørholm, Morten H. H. Jensen, Kenneth |
author_sort | Heyde, Sophia A. H. |
collection | PubMed |
description | Poly(ethylene terephthalate) (PET) is the world’s most abundant polyester plastic, and its ongoing accumulation in nature is causing a global environmental problem. Currently, the main recycling processes utilize thermomechanical or chemical means, resulting in the deterioration of the mechanical properties of PET. Consequently, polluting de novo synthesis remains preferred, creating the need for more efficient and bio-sustainable ways to hydrolyze the polymer. Recently, a PETase enzyme from the bacterium Ideonella sakaiensis was shown to facilitate PET biodegradation, albeit at slow rate. Engineering of more efficient PETases is required for industrial relevance, but progress is currently hampered by the dependency on intracellular expression in Escherichia coli. To create a more efficient screening platform in E. coli, we explore different surface display anchors for fast and easy assaying of PETase activity. We show that PETases can be functionally displayed on the bacterial cell surface, enabling screening of enzyme activity on PET microparticles – both while anchored to the cell and following solubilization of the enzymes. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-021-01582-7. |
format | Online Article Text |
id | pubmed-8088578 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-80885782021-05-03 Surface display as a functional screening platform for detecting enzymes active on PET Heyde, Sophia A. H. Arnling Bååth, Jenny Westh, Peter Nørholm, Morten H. H. Jensen, Kenneth Microb Cell Fact Research Poly(ethylene terephthalate) (PET) is the world’s most abundant polyester plastic, and its ongoing accumulation in nature is causing a global environmental problem. Currently, the main recycling processes utilize thermomechanical or chemical means, resulting in the deterioration of the mechanical properties of PET. Consequently, polluting de novo synthesis remains preferred, creating the need for more efficient and bio-sustainable ways to hydrolyze the polymer. Recently, a PETase enzyme from the bacterium Ideonella sakaiensis was shown to facilitate PET biodegradation, albeit at slow rate. Engineering of more efficient PETases is required for industrial relevance, but progress is currently hampered by the dependency on intracellular expression in Escherichia coli. To create a more efficient screening platform in E. coli, we explore different surface display anchors for fast and easy assaying of PETase activity. We show that PETases can be functionally displayed on the bacterial cell surface, enabling screening of enzyme activity on PET microparticles – both while anchored to the cell and following solubilization of the enzymes. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-021-01582-7. BioMed Central 2021-05-01 /pmc/articles/PMC8088578/ /pubmed/33933097 http://dx.doi.org/10.1186/s12934-021-01582-7 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Heyde, Sophia A. H. Arnling Bååth, Jenny Westh, Peter Nørholm, Morten H. H. Jensen, Kenneth Surface display as a functional screening platform for detecting enzymes active on PET |
title | Surface display as a functional screening platform for detecting enzymes active on PET |
title_full | Surface display as a functional screening platform for detecting enzymes active on PET |
title_fullStr | Surface display as a functional screening platform for detecting enzymes active on PET |
title_full_unstemmed | Surface display as a functional screening platform for detecting enzymes active on PET |
title_short | Surface display as a functional screening platform for detecting enzymes active on PET |
title_sort | surface display as a functional screening platform for detecting enzymes active on pet |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8088578/ https://www.ncbi.nlm.nih.gov/pubmed/33933097 http://dx.doi.org/10.1186/s12934-021-01582-7 |
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