Rational Protein Engineering to Increase the Activity and Stability of <i>Is</i>PETase Using the PROSS Algorithm

Polyethylene terephthalate (PET) is the most widely used polyester plastic, with applications in the textile and packaging industry. Currently, re-moulding is the main path for PET recycling, but this eventually leads to an unsustainable loss of quality; thus, other means of recycling are required....

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Autores principales:	Andrew Rennison, Jakob R. Winther, Cristiano Varrone
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	PETase thermostability thermal deactivation assay PET hydrolysis Organic chemistry QD241-441
Acceso en línea:	https://doaj.org/article/77ae5989fdbd43d8a835a712e214b01e
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spelling	oai:doaj.org-article:77ae5989fdbd43d8a835a712e214b01e2021-11-25T18:47:54ZRational Protein Engineering to Increase the Activity and Stability of <i>Is</i>PETase Using the PROSS Algorithm10.3390/polym132238842073-4360https://doaj.org/article/77ae5989fdbd43d8a835a712e214b01e2021-11-01T00:00:00Zhttps://www.mdpi.com/2073-4360/13/22/3884https://doaj.org/toc/2073-4360Polyethylene terephthalate (PET) is the most widely used polyester plastic, with applications in the textile and packaging industry. Currently, re-moulding is the main path for PET recycling, but this eventually leads to an unsustainable loss of quality; thus, other means of recycling are required. Enzymatic hydrolysis offers the possibility of monomer formation under mild conditions and opens up alternative and infinite recycling paths. Here, <i>Is</i>PETase, derived from the bacterium <i>Ideonella sakaiensis</i>, is considered to be the most active enzyme for PET degradation under mild conditions, and although several studies have demonstrated improvements to both the stability and activity of this enzyme, stability at even moderate temperatures is still an issue. In the present study, we have used sequence and structure-based bioinformatic tools to identify mutations to increase the thermal stability of the enzyme so as to increase PET degradation activity during extended hydrolysis reactions. We found that amino acid substitution S136E showed significant increases to activity and stability. S136E is a previously unreported variant that led to a 3.3-fold increase in activity relative to wild type.Andrew RennisonJakob R. WintherCristiano VarroneMDPI AGarticlePETasethermostabilitythermal deactivation assayPET hydrolysisOrganic chemistryQD241-441ENPolymers, Vol 13, Iss 3884, p 3884 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	PETase thermostability thermal deactivation assay PET hydrolysis Organic chemistry QD241-441
spellingShingle	PETase thermostability thermal deactivation assay PET hydrolysis Organic chemistry QD241-441 Andrew Rennison Jakob R. Winther Cristiano Varrone Rational Protein Engineering to Increase the Activity and Stability of <i>Is</i>PETase Using the PROSS Algorithm
description	Polyethylene terephthalate (PET) is the most widely used polyester plastic, with applications in the textile and packaging industry. Currently, re-moulding is the main path for PET recycling, but this eventually leads to an unsustainable loss of quality; thus, other means of recycling are required. Enzymatic hydrolysis offers the possibility of monomer formation under mild conditions and opens up alternative and infinite recycling paths. Here, <i>Is</i>PETase, derived from the bacterium <i>Ideonella sakaiensis</i>, is considered to be the most active enzyme for PET degradation under mild conditions, and although several studies have demonstrated improvements to both the stability and activity of this enzyme, stability at even moderate temperatures is still an issue. In the present study, we have used sequence and structure-based bioinformatic tools to identify mutations to increase the thermal stability of the enzyme so as to increase PET degradation activity during extended hydrolysis reactions. We found that amino acid substitution S136E showed significant increases to activity and stability. S136E is a previously unreported variant that led to a 3.3-fold increase in activity relative to wild type.
format	article
author	Andrew Rennison Jakob R. Winther Cristiano Varrone
author_facet	Andrew Rennison Jakob R. Winther Cristiano Varrone
author_sort	Andrew Rennison
title	Rational Protein Engineering to Increase the Activity and Stability of <i>Is</i>PETase Using the PROSS Algorithm
title_short	Rational Protein Engineering to Increase the Activity and Stability of <i>Is</i>PETase Using the PROSS Algorithm
title_full	Rational Protein Engineering to Increase the Activity and Stability of <i>Is</i>PETase Using the PROSS Algorithm
title_fullStr	Rational Protein Engineering to Increase the Activity and Stability of <i>Is</i>PETase Using the PROSS Algorithm
title_full_unstemmed	Rational Protein Engineering to Increase the Activity and Stability of <i>Is</i>PETase Using the PROSS Algorithm
title_sort	rational protein engineering to increase the activity and stability of <i>is</i>petase using the pross algorithm
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/77ae5989fdbd43d8a835a712e214b01e
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Rational Protein Engineering to Increase the Activity and Stability of <i>Is</i>PETase Using the PROSS Algorithm

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