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Development of Clostridium saccharoperbutylacetonicum as a Whole Cell Biocatalyst for Production of Chirally Pure (R)-1,3-Butanediol

Grosse-Honebrink, Alexander; Little, Gareth T.; Bean, Zak; Heldt, Dana; Cornock, Ruth H. M.; Winzer, Klaus; Minton, Nigel P.; Green, Edward; Zhang, Ying

Development of Clostridium saccharoperbutylacetonicum as a Whole Cell Biocatalyst for Production of Chirally Pure (R)-1,3-Butanediol Thumbnail


Authors

Alexander Grosse-Honebrink

Gareth T. Little

Zak Bean

Dana Heldt

Ruth H. M. Cornock

Edward Green

YING ZHANG YING.ZHANG@NOTTINGHAM.AC.UK
Assistant Professor



Abstract

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.

Citation

Grosse-Honebrink, A., Little, G. T., Bean, Z., Heldt, D., Cornock, R. H. M., Winzer, K., …Zhang, Y. (2021). Development of Clostridium saccharoperbutylacetonicum as a Whole Cell Biocatalyst for Production of Chirally Pure (R)-1,3-Butanediol. Frontiers in Bioengineering and Biotechnology, 9, Article 659895. https://doi.org/10.3389/fbioe.2021.659895

Journal Article Type Article
Acceptance Date Apr 12, 2021
Online Publication Date May 13, 2021
Publication Date May 13, 2021
Deposit Date Apr 28, 2021
Publicly Available Date May 13, 2021
Journal Frontiers in Bioengineering and Biotechnology
Electronic ISSN 2296-4185
Publisher Frontiers Media
Peer Reviewed Peer Reviewed
Volume 9
Article Number 659895
DOI https://doi.org/10.3389/fbioe.2021.659895
Keywords (R)-1; 3-butanediol; Clostridium saccharoperbutylacetonicum; biotechnology; phaB; 7 allele-coupled exchange; codon optimization; enantiomeric excess; biocatalyst 8
Public URL https://nottingham-repository.worktribe.com/output/5500909
Publisher URL https://www.frontiersin.org/articles/10.3389/fbioe.2021.659895/full

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