Development of Clostridium saccharoperbutylacetonicum as a Whole Cell Biocatalyst for Production of Chirally Pure (R)-1,3-Butanediol
Chirally pure (R)-1,3-butanediol ((R)-1,3-BDO) is a valuable intermediate for the production of fragrances, pheromones, insecticides and antibiotics. Biotechnological production results in superior enantiomeric excess over chemical production and is therefore the preferred production route. In this...
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doaj-cb18d2e997ad41de910ee6307f645d372021-05-13T09:48:23ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852021-05-01910.3389/fbioe.2021.659895659895Development of Clostridium saccharoperbutylacetonicum as a Whole Cell Biocatalyst for Production of Chirally Pure (R)-1,3-ButanediolAlexander Grosse-Honebrink0Gareth T. Little1Zak Bean2Dana Heldt3Ruth H. M. Cornock4Klaus Winzer5Nigel P. Minton6Edward Green7Ying Zhang8Clostridia Research Group, BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), Biodiscovery Institute, School of Life Sciences, University of Nottingham, Nottingham, United KingdomClostridia Research Group, BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), Biodiscovery Institute, School of Life Sciences, University of Nottingham, Nottingham, United KingdomCHAIN Biotechnology Ltd., MediCity, Nottingham, United KingdomCHAIN Biotechnology Ltd., MediCity, Nottingham, United KingdomClostridia Research Group, BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), Biodiscovery Institute, School of Life Sciences, University of Nottingham, Nottingham, United KingdomClostridia Research Group, BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), Biodiscovery Institute, School of Life Sciences, University of Nottingham, Nottingham, United KingdomClostridia Research Group, BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), Biodiscovery Institute, School of Life Sciences, University of Nottingham, Nottingham, United KingdomCHAIN Biotechnology Ltd., MediCity, Nottingham, United KingdomClostridia Research Group, BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), Biodiscovery Institute, School of Life Sciences, University of Nottingham, Nottingham, United KingdomChirally pure (R)-1,3-butanediol ((R)-1,3-BDO) is a valuable intermediate for the production of fragrances, pheromones, insecticides and antibiotics. Biotechnological production results in superior enantiomeric excess over chemical production and is therefore the preferred production route. In this study (R)-1,3-BDO was produced in the industrially important whole cell biocatalyst Clostridium saccharoperbutylacetonicum through expression of the enantio-specific phaB gene from Cupriavidus necator. The heterologous pathway was optimised in three ways: at the transcriptional level choosing strongly expressed promoters and comparing plasmid borne with chromosomal gene expression, at the translational level by optimising the codon usage of the gene to fit the inherent codon adaptation index of C. saccharoperbutylacetonicum, and at the enzyme level by introducing point mutations which led to increased enzymatic activity. The resulting whole cell catalyst produced up to 20 mM (1.8 g/l) (R)-1,3-BDO in non-optimised batch fermentation which is a promising starting position for economical production of this chiral chemical.https://www.frontiersin.org/articles/10.3389/fbioe.2021.659895/fullClostridium saccharoperbutylacetonicumbiotechnologyphaBAllele Coupled Exchange(R)-13-butanediol |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Alexander Grosse-Honebrink Gareth T. Little Zak Bean Dana Heldt Ruth H. M. Cornock Klaus Winzer Nigel P. Minton Edward Green Ying Zhang |
spellingShingle |
Alexander Grosse-Honebrink Gareth T. Little Zak Bean Dana Heldt Ruth H. M. Cornock Klaus Winzer Nigel P. Minton Edward Green Ying Zhang Development of Clostridium saccharoperbutylacetonicum as a Whole Cell Biocatalyst for Production of Chirally Pure (R)-1,3-Butanediol Frontiers in Bioengineering and Biotechnology Clostridium saccharoperbutylacetonicum biotechnology phaB Allele Coupled Exchange (R)-1 3-butanediol |
author_facet |
Alexander Grosse-Honebrink Gareth T. Little Zak Bean Dana Heldt Ruth H. M. Cornock Klaus Winzer Nigel P. Minton Edward Green Ying Zhang |
author_sort |
Alexander Grosse-Honebrink |
title |
Development of Clostridium saccharoperbutylacetonicum as a Whole Cell Biocatalyst for Production of Chirally Pure (R)-1,3-Butanediol |
title_short |
Development of Clostridium saccharoperbutylacetonicum as a Whole Cell Biocatalyst for Production of Chirally Pure (R)-1,3-Butanediol |
title_full |
Development of Clostridium saccharoperbutylacetonicum as a Whole Cell Biocatalyst for Production of Chirally Pure (R)-1,3-Butanediol |
title_fullStr |
Development of Clostridium saccharoperbutylacetonicum as a Whole Cell Biocatalyst for Production of Chirally Pure (R)-1,3-Butanediol |
title_full_unstemmed |
Development of Clostridium saccharoperbutylacetonicum as a Whole Cell Biocatalyst for Production of Chirally Pure (R)-1,3-Butanediol |
title_sort |
development of clostridium saccharoperbutylacetonicum as a whole cell biocatalyst for production of chirally pure (r)-1,3-butanediol |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Bioengineering and Biotechnology |
issn |
2296-4185 |
publishDate |
2021-05-01 |
description |
Chirally pure (R)-1,3-butanediol ((R)-1,3-BDO) is a valuable intermediate for the production of fragrances, pheromones, insecticides and antibiotics. Biotechnological production results in superior enantiomeric excess over chemical production and is therefore the preferred production route. In this study (R)-1,3-BDO was produced in the industrially important whole cell biocatalyst Clostridium saccharoperbutylacetonicum through expression of the enantio-specific phaB gene from Cupriavidus necator. The heterologous pathway was optimised in three ways: at the transcriptional level choosing strongly expressed promoters and comparing plasmid borne with chromosomal gene expression, at the translational level by optimising the codon usage of the gene to fit the inherent codon adaptation index of C. saccharoperbutylacetonicum, and at the enzyme level by introducing point mutations which led to increased enzymatic activity. The resulting whole cell catalyst produced up to 20 mM (1.8 g/l) (R)-1,3-BDO in non-optimised batch fermentation which is a promising starting position for economical production of this chiral chemical. |
topic |
Clostridium saccharoperbutylacetonicum biotechnology phaB Allele Coupled Exchange (R)-1 3-butanediol |
url |
https://www.frontiersin.org/articles/10.3389/fbioe.2021.659895/full |
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