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| Improved Production of 2,3-Butanediol by Enhancing the Level of Intracellular NADH and Activity of Acetoin Reductase |
| HU Gui-yuan, YANG Tao-wei, RAO Zhi-ming, LIU Mei, XU Mei-juan, ZHANG Xian |
| Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China |
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Abstract Acetoin is the main by-product during the 2,3-butanediol(2,3-BD) fermentation. Acetoin reductase (ACR) catalyzes the conversion of acetoin to 2,3-BD with concomitant oxidation of NADH to NAD+. Therefore, intracellular 2,3-BD production is likely governed by the quantities of rate-limiting factor(s) ACR and/or NADH. In bacterial metabolism, glucose is first converted to pyruvate before generation of major products, then pyruvate produced from glucose is channeled into 2,3-BD fermentation pathway. While, glucose can be converted to pyruvate through two main metabolism pathway PPP and EMP. However, there are NADPH generated via the former pathway, which was not necessary for 2,3-BD production. Glucose-6-phosphate dehydrogenase(G6PDH) is the key enzyme in PPP pathway and to enhance the level of intracellular NADH, metabolic flux of EMP pathway was increased by disrupting G6PDH, an key enzyme in PPP pathway. Flask fermentation experiments indicates that the level of intracellular NADH was enhanced in the recombinant strain B. subtilis168△zwf, and the yield of 2,3-BD was increased by 15.0%, and the titer of main by-products AC was decreased by 10.6%, however, the accumulation of acetic acid, lactic acid and other organic acid were enhanced. In order to improve the yield and production efficiency of 2,3-BD further, a recombinant strain B.subtilis168△zwf/pMA5-kphs was constructed by overexpressing ACR in B.subtilis168△zwf. Compared with parental strain, the yield of 2,3-BD was increased by 37.3 %, AC was decreased by 28.1%, and acetate acid, lactate acid, succinic acid and other by-products also were decreased.
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Received: 05 January 2016
Published: 15 February 2016
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