| 过量表达烟酸转磷酸核糖激酶对大肠杆菌NZN111产丁二酸的影响 |
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| 引用本文： | 刘嵘明,马江锋,梁丽亚,徐冰,王光明,张敏,姜岷.过量表达烟酸转磷酸核糖激酶对大肠杆菌NZN111产丁二酸的影响[J].生物工程学报,2011,27(10):1438-1447. |
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| 作者姓名： | 刘嵘明 马江锋 梁丽亚 徐冰 王光明 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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| 摘 要： | 大肠杆菌NZN111厌氧发酵的主要产物为丁二酸，是发酵生产丁二酸的潜力菌株。但是由于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)，导致辅酶NADH/NAD+不平衡，厌氧条件下不能利用葡萄糖生长代谢。构建烟酸转磷酸核糖激酶的重组菌Escherichia coli NZN111/pTrc99a-pncB，在厌氧摇瓶发酵过程中通过添加0.5 mmol/L的烟酸、0.3 mmol/L的IPTG诱导后重组菌的烟酸转磷酸核糖激酶 (Nicotinic acid phosphor
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| 关 键 词： | 烟酸转磷酸核糖激酶，大肠杆菌NZN111，丁二酸，两阶段发酵 |
| 收稿时间： | 2011/2/16 0:00:00 |
Effect of overexpression of nicotinic acid phosphoribosyl transferase on succinic acid production in Escherichia coli NZN111 |
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| Rongming Liu,Jiangfeng M,Liya Liang,Bing Xu,Guangming Wang,Min Zhang and Min Jiang.Effect of overexpression of nicotinic acid phosphoribosyl transferase on succinic acid production in Escherichia coli NZN111[J].Chinese Journal of Biotechnology,2011,27(10):1438-1447. |
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| Authors: | Rongming Liu Jiangfeng M Liya Liang Bing Xu Guangming Wang 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 strain NZN111 is a promising candidate for the fermentative production of succinate. However, because lactate dehydrogenase and pyruvate formate lyase were inactivated in NZN111, this strain had an unbalanced NADH/NAD+ ratio and could not use glucose under anaerobic conditions. In this study, a recombinant strain E. coli NZN111/pTrc99a-pncB was constructed to overexpress the nicotinic acid phosphoribosyl transferase gene (pncB). Under anaerobic conditions with the addition of 0.5 mmol/L nicotinic acid and 0.3 mmol/L isopropyl beta-D-thiogalactopyranoside (IPTG), the specific nicotinic acid phosphoribosyl transferase (NAPRTase, EC 2.4.2.11) activity in the recombinant strain was 11-fold higher than that in E. coli NZN111, the concentration of NAD(H) was increased by 3.85-fold, especially the concentration of NAD+ was increased by 5.17-fold and NADH/NAD+ was decreased from 0.640 to 0.125. The recombinant strain regained the capability of growth and glucose utilization under anaerobic conditions. |
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| Keywords: | nicotinic acid phosphoribosyl transferase Escherichia coli NZN111 succinic acid dual-phase fermentation |
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