Journal
CATALYSTS
Volume 11, Issue 12, Pages -Publisher
MDPI
DOI: 10.3390/catal11121422
Keywords
2; 3-butanediol; acetoin; Serratia marcescens; whole-cell biocatalysts; NADH; NAD(+) regeneration system
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Funding
- Leading Talent Project of the Hubei Academy of Agricultural Sciences [L2018030]
- Wuhan Science and Technology Plan Project [2018020401011305]
- China National Key Research and Development (RD) Program [2021YFC2100100]
- Open Funding Project of the State Key Laboratory of Biocatalysis and Enzyme Engineering [SKLBEE2018003, 202001BA070001-086]
- Hubei Provincial Central Government Guides Local Science and Technology Development Special Project [2018ZYYD015]
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An auto-inducing expression system was developed to express target genes in S. marcescens MG1, creating a whole-cell biocatalyst for 2,3-butanediol and acetoin production. Enzymes were co-expressed to build NADH regeneration systems for transforming diacetyl to 2,3-butanediol and acetoin respectively, showcasing high productivities and activities in the engineered strains. This study demonstrates the potential of using S. marcescens as an effective whole-cell biocatalyst for bio-based chemical production.
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.
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