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Biochemical Characterization and NMR Study of a PET-Hydrolyzing Cutinase from Fusarium solani pisi

[Image: see text] In recent years, the drawbacks of plastics have become evident, with plastic pollution becoming a major environmental issue. There is an urgent need to find solutions to efficiently manage plastic waste by using novel recycling methods. Biocatalytic recycling of plastics by using e...

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Autores principales: Hellesnes, Kristina Naasen, Vijayaraj, Shunmathi, Fojan, Peter, Petersen, Evamaria, Courtade, Gaston
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2023
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10116592/
https://www.ncbi.nlm.nih.gov/pubmed/36967526
http://dx.doi.org/10.1021/acs.biochem.2c00619
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author Hellesnes, Kristina Naasen
Vijayaraj, Shunmathi
Fojan, Peter
Petersen, Evamaria
Courtade, Gaston
author_facet Hellesnes, Kristina Naasen
Vijayaraj, Shunmathi
Fojan, Peter
Petersen, Evamaria
Courtade, Gaston
author_sort Hellesnes, Kristina Naasen
collection PubMed
description [Image: see text] In recent years, the drawbacks of plastics have become evident, with plastic pollution becoming a major environmental issue. There is an urgent need to find solutions to efficiently manage plastic waste by using novel recycling methods. Biocatalytic recycling of plastics by using enzyme-catalyzed hydrolysis is one such solution that has gained interest, in particular for recycling poly(ethylene terephthalate) (PET). To provide insights into PET hydrolysis by cutinases, we have here characterized the kinetics of a PET-hydrolyzing cutinase from Fusarium solani pisi (FsC) at different pH values, mapped the interaction between FsC and the PET analogue BHET by using NMR spectroscopy, and monitored product release directly and in real time by using time-resolved NMR experiments. We found that primarily aliphatic side chains around the active site participate in the interaction with BHET and that pH conditions and a mutation around the active site (L182A) can be used to tune the relative amounts of degradation products. Moreover, we propose that the low catalytic performance of FsC on PET is caused by poor substrate binding combined with slow MHET hydrolysis. Overall, our results provide insights into obstacles that preclude efficient PET hydrolysis by FsC and suggest future approaches for overcoming these obstacles and generating efficient PET-hydrolyzing enzymes.
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spelling pubmed-101165922023-04-21 Biochemical Characterization and NMR Study of a PET-Hydrolyzing Cutinase from Fusarium solani pisi Hellesnes, Kristina Naasen Vijayaraj, Shunmathi Fojan, Peter Petersen, Evamaria Courtade, Gaston Biochemistry [Image: see text] In recent years, the drawbacks of plastics have become evident, with plastic pollution becoming a major environmental issue. There is an urgent need to find solutions to efficiently manage plastic waste by using novel recycling methods. Biocatalytic recycling of plastics by using enzyme-catalyzed hydrolysis is one such solution that has gained interest, in particular for recycling poly(ethylene terephthalate) (PET). To provide insights into PET hydrolysis by cutinases, we have here characterized the kinetics of a PET-hydrolyzing cutinase from Fusarium solani pisi (FsC) at different pH values, mapped the interaction between FsC and the PET analogue BHET by using NMR spectroscopy, and monitored product release directly and in real time by using time-resolved NMR experiments. We found that primarily aliphatic side chains around the active site participate in the interaction with BHET and that pH conditions and a mutation around the active site (L182A) can be used to tune the relative amounts of degradation products. Moreover, we propose that the low catalytic performance of FsC on PET is caused by poor substrate binding combined with slow MHET hydrolysis. Overall, our results provide insights into obstacles that preclude efficient PET hydrolysis by FsC and suggest future approaches for overcoming these obstacles and generating efficient PET-hydrolyzing enzymes. American Chemical Society 2023-03-27 /pmc/articles/PMC10116592/ /pubmed/36967526 http://dx.doi.org/10.1021/acs.biochem.2c00619 Text en © 2023 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Hellesnes, Kristina Naasen
Vijayaraj, Shunmathi
Fojan, Peter
Petersen, Evamaria
Courtade, Gaston
Biochemical Characterization and NMR Study of a PET-Hydrolyzing Cutinase from Fusarium solani pisi
title Biochemical Characterization and NMR Study of a PET-Hydrolyzing Cutinase from Fusarium solani pisi
title_full Biochemical Characterization and NMR Study of a PET-Hydrolyzing Cutinase from Fusarium solani pisi
title_fullStr Biochemical Characterization and NMR Study of a PET-Hydrolyzing Cutinase from Fusarium solani pisi
title_full_unstemmed Biochemical Characterization and NMR Study of a PET-Hydrolyzing Cutinase from Fusarium solani pisi
title_short Biochemical Characterization and NMR Study of a PET-Hydrolyzing Cutinase from Fusarium solani pisi
title_sort biochemical characterization and nmr study of a pet-hydrolyzing cutinase from fusarium solani pisi
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10116592/
https://www.ncbi.nlm.nih.gov/pubmed/36967526
http://dx.doi.org/10.1021/acs.biochem.2c00619
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