Efficient production of 2,3-butanediol in Saccharomyces cerevisiae by eliminating ethanol and glycerol production and redox rebalancing |
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| Affiliation: | 1. State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People׳s Republic of China;2. State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University, Shanghai 200240, People׳s Republic of China;1. Department of Agricultural Biotechnology, Seoul National University, Seoul 151-921, Republic of Korea;2. Center for Food and Bioconvergence, Seoul National University, Seoul 151-921, Republic of Korea;3. Department of Bio and Fermentation Convergence Technology, Kookmin University, Seoul 136-702, Republic of Korea;1. State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University, Shanghai 200240, PR China;2. State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, PR China;1. Key Laboratory of Environment Correlative Dietology (Ministry of Education), College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China;2. Hubei Collaborative Innovation Center for Green Transformation of Bio-Resources, College of Life Sciences, Hubei University, Wuhan 430062, China;3. State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China;4. Department of Food Science and Human Nutrition, Iowa State University, 50011 Ames, IA, USA;1. Institute of Microbiology, Bulgarian Academy of Sciences, Bl. 26 Acad. G. Bonchev Str., 1113 Sofia, Bulgaria;2. Institute of Chemical Engineering, Bulgarian Academy of Sciences, Bl. 103 Acad. G. Bonchev Str., 1113 Sofia, Bulgaria;1. Bioprocessing and Renewable Energy Laboratory, Department of Grain Science and Industry, Kansas State University, Manhattan, KS 66506, USA;2. Department of Chemical Engineering, Kansas State University, Manhattan, KS 66506, USA |
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| Abstract: | 2,3-Butanediol is a promising valuable chemical that can be used in various areas as a liquid fuel and a platform chemical. Here, 2,3-butanediol production in Saccharomyces cerevisiae was improved stepwise by eliminating byproduct formation and redox rebalancing. By introducing heterologous 2,3-butanediol biosynthetic pathway and deleting competing pathways producing ethanol and glycerol, metabolic flux was successfully redirected to 2,3-butanediol. In addition, the resulting redox cofactor imbalance was restored by overexpressing water-forming NADH oxidase (NoxE) from Lactococcus lactis. In a flask fed-batch fermentation with optimized conditions, the engineered adh1Δadh2Δadh3Δadh4Δadh5Δgpd1Δgpd2Δ strain overexpressing Bacillus subtilis α-acetolactate synthase (AlsS) and α-acetolactate decarboxylase (AlsD), S. cerevisiae 2,3-butanediol dehydrogenase (Bdh1), and L. lactis NoxE from a single multigene-expression vector produced 72.9 g/L 2,3-butanediol with the highest yield (0.41 g/g glucose) and productivity (1.43 g/(L·h)) ever reported in S. cerevisiae. |
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| Keywords: | Alcohol dehydrogenase 2,3-Butanediol Glycerol-3-phosphate dehydrogenase NADH oxidase |
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