Enhancing the carbon flux and NADPH supply to increase L-isoleucine production in Corynebacterium glutamicum
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- 作者：Lianghong Yin (1) (2)
Jianxun Zhao (1) (2)
Cheng Chen (1) (2)
Xiaoqing Hu (1)
Xiaoyuan Wang (1) (2) - 关键词：Corynebacterium glutamicum ; L ; isoleucine ; ilvBN ; ilvA ; ppnk ; lrp ; brnFE ; NADPH
- 刊名：Biotechnology and Bioprocess Engineering
- 出版年：2014
- 出版时间：February 2014
- 年：2014
- 卷：19
- 期：1
- 页码：132-142
- 全文大小：486 KB
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Jianxun Zhao (1) (2)
Cheng Chen (1) (2)
Xiaoqing Hu (1)
Xiaoyuan Wang (1) (2)
1. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214-122, China
2. Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214-122, China - ISSN：1976-3816
文摘
Our previous work has shown that L-isoleucine production in Corynebacterium glutamicum IWJ001 could be increased by overexpressing ilvA1 encoding a feedback-resistant threonine dehydratase, ilvBN1 encoding a feedback-resistant acetohydroxy acid synthase, lrp encoding the global regulator Lrp, brnFE encoding the two-component export system BrnFE, or ppnk1 encoding NAD kinase. The main purpose of this study is to further increase the L-isoleucine production in C. glutamicum IWJ001 by overexpressing the above genes in various combinations. Several C. glutamicum strains IWJ001/pDXW-8-ppnk1-lrp-brnFE, IWJ001/pDXW-8-ilvBN1-ilvA1-lrp-brnFE, IWJ001/pDXW-8-ilvBN1-ilvA1-ppnk1, and IWJ001/pDXW-8-ppnk1-ilvBN1-ilvA1-lrp-brnFE were constructed, and L-isoleucine production and activities of several key enzymes in these strains were analyzed. Compared with the control strain IWJ001/pDXW-8, L-isoleucine production increased in all of the four strains. IWJ001/pDXW-8-ilvBN1-ilvA1-ppnk1 showed the highest L-isoleucine production and produced 32.3 g/L L-isoleucine in 72 h fed batch fermentation. The results indicate that L-isoleucine production in C. glutamicum could be increased by enhancing the carbon flux and NADPH supply in the biosynthetic pathway.