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食品与发酵工业  2019, Vol. 45 Issue (10): 1-9    DOI: 10.13995/j.cnki.11-1802/ts.019690
  研究报告 本期目录 | 过刊浏览 | 高级检索 |
阻断辅因子NADPH合成对谷氨酸棒杆菌生长及产物合成的影响
杨汉昆1,2, 徐建中1,2*, 张伟国1,2*
1(江南大学 生物工程学院,江苏 无锡,214122)
2(工业生物技术教育部重点实验室(江南大学),江苏 无锡,214122)
Blocking NADPH biosynthesis affected growth and metabolites formation of Corynebacterium glutamicum
YANG Hankun1,2, XU Jianzhong1,2*, ZHANG Weiguo1,2*
1(School of Biotechnology, Jiangnan University,Wuxi 214122,China)
2(Key Laboratory of Industrial Biotechnology, Ministry of Education (Jiangnan University),Wuxi 214122,China)
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摘要 该文通过敲除谷氨酸棒杆菌(C. glutamicum)中参与NADPH合成的葡萄糖-6-磷酸脱氢酶基因(zwf)、苹果酸酶基因(malE),并将自身NADP+-依赖型异柠檬酸脱氢酶基因(icdCg)替换为变形链球菌(Streptococcus mutans)NAD+-依赖型异柠檬酸脱氢酶基因(icdSm),构建重组菌株C. glutamicum LYS Δzwf ΔmalE ΔicdCg::icdSm,并研究阻断NADPH合成途径对胞内腺嘌呤核苷酸、吡啶核苷酸、细胞生长、葡萄糖消耗速率、产物合成的影响。与出发菌C. glutamicum LYS相比,重组菌胞内NADH提高38.58%,NADH/NAD+提高53.84%。而NADPH降低89.51%,明显低于出发菌,NADPH/NADP+降低93.13%。摇瓶结果表明,重组菌葡萄糖代谢能力明显减弱,生长也相对缓慢,菌体量降低4.36%。同时L-赖氨酸产量降低87.69%,而在胞内积累了大量副产物,如丙酮酸、乳酸等。为进一步研究NADPH调控L-赖氨酸产生菌胞内微环境的生理机制提供了研究基础。
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杨汉昆
徐建中
张伟国
关键词:  谷氨酸棒杆菌  还原型烟酰胺腺嘌呤二核苷酸磷酸  L-赖氨酸  代谢物    
Abstract: NADPH plays important roles in regulating cell growth and L-lysine synthesis in Corynebacterium glutamicum. A recombinant strain with blocked NADPH biosynthesis was constructed by deleting the genes of glucose-6-phosphate dehydrogenase (zwf) and malic enzyme (malE) and by replacing NADP+-dependent isocitrate dehydrogenase gene (icdCg) with Streptococcus mutans NAD+-dependent isocitrate dehydrogenase gene (icdSm). Compared with C. glutamicum LYS, the levels of intracellular NADH and NADH/NAD+ in the recombinant strain increased by 38.58% and 53.84%, respectively, while the intracellular NADPH significantly decreased by 89.51%, and intracellular NADPH/NADP+ decreased by 93.13%. Meanwhile, the recombinant strain showed slower cell growth and its glucose metabolism significantly lowered during shake-flask fermentation. As a result, the biomass reduced 4.36% and the yield of L-lysine decreased by 87.69%, together with more by-products (pyruvate and lactate etc.) accumulated in cells. This study provides a basis for further studies on physiological mechanisms of NADPH that regulates intracellular microenvironment of L-lysine producing strains.
Key words:  Corynebacterium glutamicum    NADPH    L-lysine    metabolites
收稿日期:  2018-12-20                出版日期:  2019-05-25      发布日期:  2019-06-17      期的出版日期:  2019-05-25
基金资助: 国家自然科学基金青年基金项目(31601459);江苏高校品牌专业建设工程资助项目
作者简介:  硕士研究生(张伟国教授和徐建中副教授为共同通讯作者,E-mail:zhangwg168@126.com,xujz126@126.com)。
引用本文:    
杨汉昆,徐建中,张伟国. 阻断辅因子NADPH合成对谷氨酸棒杆菌生长及产物合成的影响[J]. 食品与发酵工业, 2019, 45(10): 1-9.
YANG Hankun,XU Jianzhong,ZHANG Weiguo. Blocking NADPH biosynthesis affected growth and metabolites formation of Corynebacterium glutamicum[J]. Food and Fermentation Industries, 2019, 45(10): 1-9.
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http://sf1970.cnif.cn/CN/10.13995/j.cnki.11-1802/ts.019690  或          http://sf1970.cnif.cn/CN/Y2019/V45/I10/1
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