| 过量表达苹果酸脱氢酶对大肠杆菌NZN111产丁二酸的影响 |
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| 引用本文： | 梁丽亚,马江锋,刘嵘明,王光明,徐冰,张敏,姜岷.过量表达苹果酸脱氢酶对大肠杆菌NZN111产丁二酸的影响[J].生物工程学报,2011,27(7):1005-1012. |
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| 作者姓名： | 梁丽亚 马江锋 刘嵘明 王光明 徐冰 张敏 姜岷 |
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| 作者单位： | 南京工业大学生物与制药工程学院材料化学工程国家重点实验室，南京，210009 |
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| 基金项目： | 国家自然科学基金 (No. 21076105)，国家重点基础研究发展计划 (973计划) (No. 2009CB724701) 资助。 |
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| 摘 要： | 大肠杆菌NZN111是敲除了乳酸脱氢酶的编码基因 (ldhA) 和丙酮酸-甲酸裂解酶的编码基因 (pflB) 的工程菌，厌氧条件下由于辅酶NAD(H) 的不平衡导致其丧失了代谢葡萄糖的能力。构建了苹果酸脱氢酶的重组菌大肠杆菌NZN111/pTrc99a-mdh，在厌氧摇瓶发酵过程中通过0.3 mmol/L的IPTG诱导后重组菌的苹果酸脱氢酶 (Malate dehydrogenase，MDH) 酶活较出发菌株提高了14.8倍，NADH/NAD+的比例从0.64下降到0.26，同时NAD+和NADH浓度分别
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| 关 键 词： | 苹果酸脱氢酶，大肠杆菌NZN111，丁二酸，两阶段发酵 |
| 收稿时间： | 2010/12/10 0:00:00 |
Effect of overexpression of malate dehydrogenase on succinic acid production in Escherichia coli NZN111 |
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| Liya Liang,Jiangfeng M,Rongming Liu,Guangming Wang,Bing Xu,Min Zhang and Min Jiang.Effect of overexpression of malate dehydrogenase on succinic acid production in Escherichia coli NZN111[J].Chinese Journal of Biotechnology,2011,27(7):1005-1012. |
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| Authors: | Liya Liang Jiangfeng M Rongming Liu Guangming Wang Bing Xu Min Zhang and Min Jiang |
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| Institution: | State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, China;State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, China;State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, China;State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, China;State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, China;State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, China;State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, China |
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| Abstract: | Escherichia coli NZN111 is a double mutant with lactate dehydrogenase (ldhA) and pyruvate formate-lyase (pflB) inactivated. Under anaerobic conditions, disequilibrium of coenzyme NADH and NAD+ causes Escherichia coli NZN111 losing the glucose utilizing capability. In this study, we constructed a recombinant strain E. coli NZN111/pTrc99a-mdh and overexpressed the mdh gene with 0.3 mmol/L of IPTG under anaerobic fermentation condition in sealed bottles. The specific malate dehydrogenase (MDH) activity in the recombinant strain was 14.8-fold higher than that in E. coli NZN111. The NADH/ NAD+ ratio decreased from 0.64 to 0.26 and the concentration of NAD+ and NADH increased 1.5-fold and 0.2-fold respectively. Under anaerobic conditions, the recombinant strain possessed the capability of growth and glucose absorption. We took dual-phase fermentation for succinate production. After the dry cell weight (DCW) reached 6.4 g/L under aerobic conditions, the cell culture was changed to anaerobic conditions. After 15 h, 14.75 g/L glucose was consumed and succinic acid reached 15.18 g/L. The yield of succinic acid was 1.03 g/g Glu and the productivity of succinic acid was 1.012 g/(L·h). |
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| Keywords: | malate dehydrogenase Escherichia coli NZN111 succinic acid dual-phase fermentation |
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