Journal
METABOLIC ENGINEERING
Volume 23, Issue -, Pages 34-41Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2014.02.002
Keywords
Bacillus subtilis; Gene disruption; Acetoin reductase/2,3-butanediol; dehydrogenase; NADH oxidase; Acetoin pathway
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Funding
- Program for New Century Excellent Talents in University [NCET-10-0459]
- National Basic Research Program of China (973 Program) [2012CB725202]
- National Natural Science Foundation of China [21276110]
- Research Project of Chinese Ministry of Education [113033A]
- Fundamental Research Funds for the Central Universities [JUSRP51306A, JUSRP21121]
- 111 Project
- Priority Academic Program Development of Jiangsu Higher Education Institution
- [111-2-06]
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Bacillus sub tills produces acetoin as a major extracellular product. However, the by-products of 2,3-butanediol, lactic acid and ethanol were accompanied in the NADH-dependent pathways. In this work, metabolic engineering strategies were proposed to redistribute the carbon flux to acetoin by manipulation the NADH levels. We first knocked out the acetoin reductase gene bdhA to block the main flux from acetoin to 2,3-butanediol. Then, among four putative candidates, we successfully screened an active water forming NADH oxidase, YODC. Moderate expression of YODC in the bdhA disrupted B. subtilis weakened the NADH-linked pathways to by-product pools of acetoin. Through these strategies, acetoin production was improved to 56.7 g/l with an increase of 35.3%, while the production of 2,3-butanediol, lactic acid and ethanol were decreased by 92.3%, 70.1% and 75.0%, respectively, simultaneously the fermentation duration was decreased 1.7-fold. Acetoin productivity by B. subtilis was improved to 0.639 g/(1h). (C) 2014 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.
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