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Engineering of a borneol dehydrogenase from P. putida for the enzymatic resolution of camphor
ABSTRACT: Several thousand different terpenoid structures are known so far, and many of them are interesting for applications as pharmaceuticals, flavors, fragrances, biofuels, insecticides, or fine chemical intermediates. One prominent example is camphor, which has been utilized since ancient times...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Springer Berlin Heidelberg
2021
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8053192/ https://www.ncbi.nlm.nih.gov/pubmed/33846823 http://dx.doi.org/10.1007/s00253-021-11239-5 |
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author | Hofer, Michael Diener, Julia Begander, Benjamin Kourist, Robert Sieber, Volker |
author_facet | Hofer, Michael Diener, Julia Begander, Benjamin Kourist, Robert Sieber, Volker |
author_sort | Hofer, Michael |
collection | PubMed |
description | ABSTRACT: Several thousand different terpenoid structures are known so far, and many of them are interesting for applications as pharmaceuticals, flavors, fragrances, biofuels, insecticides, or fine chemical intermediates. One prominent example is camphor, which has been utilized since ancient times in medical applications. Especially (−)-camphor is gaining more and more interest for pharmaceutical applications. Hence, a commercial reliable source is needed. The natural sources for (−)-camphor are limited, and the oxidation of precious (−)-borneol would be too costly. Hence, synthesis of (−)-camphor from renewable alpha-pinene would be an inexpensive alternative. As the currently used route for the conversion of alpha-pinene to camphor produces a mixture of both enantiomers, preferably catalytic methods for the separation of this racemate are demanded to yield enantiopure camphor. Enzymatic kinetic resolution is a sustainable way to solve this challenge but requires suitable enzymes. In this study, the first borneol dehydrogenase from Pseudomonas sp. ATCC 17453, capable of catalyzing the stereoselective reduction of camphor, was examined. By using a targeted enzyme engineering approach, enantioselective enzyme variants were created with E-values > 100. The best variant was used for the enzymatic kinetic resolution of camphor racemate, yielding 79% of (−)-camphor with an ee of > 99%. KEY POINTS: • Characterization of a novel borneol dehydrogenase (BDH) from P. putida. • Development of enantioselective BDH variants for the reduction of camphor. • Enzymatic kinetic resolution of camphor with borneol dehydrogenase. GRAPHICAL ABSTRACT: [Image: see text] |
format | Online Article Text |
id | pubmed-8053192 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-80531922021-05-05 Engineering of a borneol dehydrogenase from P. putida for the enzymatic resolution of camphor Hofer, Michael Diener, Julia Begander, Benjamin Kourist, Robert Sieber, Volker Appl Microbiol Biotechnol Biotechnological Products and Process Engineering ABSTRACT: Several thousand different terpenoid structures are known so far, and many of them are interesting for applications as pharmaceuticals, flavors, fragrances, biofuels, insecticides, or fine chemical intermediates. One prominent example is camphor, which has been utilized since ancient times in medical applications. Especially (−)-camphor is gaining more and more interest for pharmaceutical applications. Hence, a commercial reliable source is needed. The natural sources for (−)-camphor are limited, and the oxidation of precious (−)-borneol would be too costly. Hence, synthesis of (−)-camphor from renewable alpha-pinene would be an inexpensive alternative. As the currently used route for the conversion of alpha-pinene to camphor produces a mixture of both enantiomers, preferably catalytic methods for the separation of this racemate are demanded to yield enantiopure camphor. Enzymatic kinetic resolution is a sustainable way to solve this challenge but requires suitable enzymes. In this study, the first borneol dehydrogenase from Pseudomonas sp. ATCC 17453, capable of catalyzing the stereoselective reduction of camphor, was examined. By using a targeted enzyme engineering approach, enantioselective enzyme variants were created with E-values > 100. The best variant was used for the enzymatic kinetic resolution of camphor racemate, yielding 79% of (−)-camphor with an ee of > 99%. KEY POINTS: • Characterization of a novel borneol dehydrogenase (BDH) from P. putida. • Development of enantioselective BDH variants for the reduction of camphor. • Enzymatic kinetic resolution of camphor with borneol dehydrogenase. GRAPHICAL ABSTRACT: [Image: see text] Springer Berlin Heidelberg 2021-04-12 2021 /pmc/articles/PMC8053192/ /pubmed/33846823 http://dx.doi.org/10.1007/s00253-021-11239-5 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This 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/) . |
spellingShingle | Biotechnological Products and Process Engineering Hofer, Michael Diener, Julia Begander, Benjamin Kourist, Robert Sieber, Volker Engineering of a borneol dehydrogenase from P. putida for the enzymatic resolution of camphor |
title | Engineering of a borneol dehydrogenase from P. putida for the enzymatic resolution of camphor |
title_full | Engineering of a borneol dehydrogenase from P. putida for the enzymatic resolution of camphor |
title_fullStr | Engineering of a borneol dehydrogenase from P. putida for the enzymatic resolution of camphor |
title_full_unstemmed | Engineering of a borneol dehydrogenase from P. putida for the enzymatic resolution of camphor |
title_short | Engineering of a borneol dehydrogenase from P. putida for the enzymatic resolution of camphor |
title_sort | engineering of a borneol dehydrogenase from p. putida for the enzymatic resolution of camphor |
topic | Biotechnological Products and Process Engineering |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8053192/ https://www.ncbi.nlm.nih.gov/pubmed/33846823 http://dx.doi.org/10.1007/s00253-021-11239-5 |
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