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Engineering of TM1459 from Thermotoga maritima for Increased Oxidative Alkene Cleavage Activity

Oxidative cleavage of alkenes is a widely employed process allowing oxyfunctionalization to corresponding carbonyl compounds. Recently, a novel biocatalytic oxidative alkene cleavage activity on styrene derivatives was identified in TM1459 from Thermotoga maritima. In this work we engineered the enz...

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Autores principales: Fink, Matthias, Trunk, Sarah, Hall, Mélanie, Schwab, Helmut, Steiner, Kerstin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5031596/
https://www.ncbi.nlm.nih.gov/pubmed/27713741
http://dx.doi.org/10.3389/fmicb.2016.01511
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author Fink, Matthias
Trunk, Sarah
Hall, Mélanie
Schwab, Helmut
Steiner, Kerstin
author_facet Fink, Matthias
Trunk, Sarah
Hall, Mélanie
Schwab, Helmut
Steiner, Kerstin
author_sort Fink, Matthias
collection PubMed
description Oxidative cleavage of alkenes is a widely employed process allowing oxyfunctionalization to corresponding carbonyl compounds. Recently, a novel biocatalytic oxidative alkene cleavage activity on styrene derivatives was identified in TM1459 from Thermotoga maritima. In this work we engineered the enzyme by site-saturation mutagenesis of active site amino acids to increase its activity and to broaden its substrate scope. A high-throughput assay for the detection of the ketone products was successfully developed. Several variants with up to twofold improved conversion level of styrene derivatives were successfully identified. Especially, changes in or removal of the C-terminus of TM1459 increased the activity most significantly. These best variants also displayed a slightly enlarged substrate scope.
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spelling pubmed-50315962016-10-06 Engineering of TM1459 from Thermotoga maritima for Increased Oxidative Alkene Cleavage Activity Fink, Matthias Trunk, Sarah Hall, Mélanie Schwab, Helmut Steiner, Kerstin Front Microbiol Microbiology Oxidative cleavage of alkenes is a widely employed process allowing oxyfunctionalization to corresponding carbonyl compounds. Recently, a novel biocatalytic oxidative alkene cleavage activity on styrene derivatives was identified in TM1459 from Thermotoga maritima. In this work we engineered the enzyme by site-saturation mutagenesis of active site amino acids to increase its activity and to broaden its substrate scope. A high-throughput assay for the detection of the ketone products was successfully developed. Several variants with up to twofold improved conversion level of styrene derivatives were successfully identified. Especially, changes in or removal of the C-terminus of TM1459 increased the activity most significantly. These best variants also displayed a slightly enlarged substrate scope. Frontiers Media S.A. 2016-09-22 /pmc/articles/PMC5031596/ /pubmed/27713741 http://dx.doi.org/10.3389/fmicb.2016.01511 Text en Copyright © 2016 Fink, Trunk, Hall, Schwab and Steiner. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Fink, Matthias
Trunk, Sarah
Hall, Mélanie
Schwab, Helmut
Steiner, Kerstin
Engineering of TM1459 from Thermotoga maritima for Increased Oxidative Alkene Cleavage Activity
title Engineering of TM1459 from Thermotoga maritima for Increased Oxidative Alkene Cleavage Activity
title_full Engineering of TM1459 from Thermotoga maritima for Increased Oxidative Alkene Cleavage Activity
title_fullStr Engineering of TM1459 from Thermotoga maritima for Increased Oxidative Alkene Cleavage Activity
title_full_unstemmed Engineering of TM1459 from Thermotoga maritima for Increased Oxidative Alkene Cleavage Activity
title_short Engineering of TM1459 from Thermotoga maritima for Increased Oxidative Alkene Cleavage Activity
title_sort engineering of tm1459 from thermotoga maritima for increased oxidative alkene cleavage activity
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5031596/
https://www.ncbi.nlm.nih.gov/pubmed/27713741
http://dx.doi.org/10.3389/fmicb.2016.01511
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