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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...

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Autores principales: Heyde, Sophia A. H., Arnling Bååth, Jenny, Westh, Peter, Nørholm, Morten H. H., Jensen, Kenneth
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
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.
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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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