Efficient 2,3-Butanediol/Acetoin Production Using Whole-Cell Biocatalyst with a New Nadh/Nad(+) Regeneration System

An auto-inducing expression system was developed that could express target genes in S. marcescens MG1. Using this system, MG1 was constructed as a whole-cell biocatalyst to produce 2,3-butanediol/acetoin. Formate dehydrogenase (FDH) and 2,3-butanediol dehydrogenase were expressed together to build a...

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Bibliographic Details
Published in:Catalysts 2021-12, Vol.11 (12), p.1422
Main Authors: Wang, Yaping, Peng, Yanhong, Liu, Xiaoyan, Zhou, Ronghua, Liao, Xianqing, Min, Yong, Hu, Yong, Wang, Ying, Rao, Ben
Format: Article
Language:English
Subjects:
2,3-butanediol
acetoin
Biocatalysts
Butanediol
Catalysts
Chemical reactions
Dehydrogenases
Fermentation
Formate dehydrogenase
Gene expression
Metabolism
NADH/NAD+ regeneration system
Nicotinamide adenine dinucleotide
Plasmids
Productivity
Proteins
Regeneration
RNA polymerase
Serratia marcescens
whole-cell biocatalysts
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Summary:An auto-inducing expression system was developed that could express target genes in S. marcescens MG1. Using this system, MG1 was constructed as a whole-cell biocatalyst to produce 2,3-butanediol/acetoin. Formate dehydrogenase (FDH) and 2,3-butanediol dehydrogenase were expressed together to build an NADH regeneration system to transform diacetyl to 2,3-butanediol. After fermentation, the extract of recombinant S. marcescens MG1ABC (pETDuet-bdhA-fdh) showed 2,3-BDH activity of 57.8 U/mg and FDH activity of 0.5 U/mg. And 27.95 g/L of 2,3-BD was achieved with a productivity of 4.66 g/Lh using engineered S. marcescens MG1(Pswnb+pETDuet-bdhA-fdh) after 6 h incubation. Next, to produce 2,3-butanediol from acetoin, NADH oxidase and 2,3-butanediol dehydrogenase from Bacillus subtilis were co-expressed to obtain a NAD+ regeneration system. After fermentation, the recombinant strain S. marcescens MG1ABC (pSWNB+pETDuet-bdhA-yodC) showed AR activity of 212.4 U/mg and NOX activity of 150.1 U/mg. We obtained 44.9 g/L of acetoin with a productivity of 3.74 g/Lh using S. marcescens MG1ABC (pSWNB+pETDuet-bdhA-yodC). This work confirmed that S. marcescens could be designed as a whole-cell biocatalyst for 2,3-butanediol and acetoin production.
ISSN:2073-4344
2073-4344
DOI:10.3390/catal11121422