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Enhancing the carbon flux and NADPH supply to increase L-isoleucine production in Corynebacterium glutamicum

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Abstract

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.

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Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214-122, China

    Lianghong Yin, Jianxun Zhao, Cheng Chen, Xiaoqing Hu & Xiaoyuan Wang

  2. Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214-122, China

    Lianghong Yin, Jianxun Zhao, Cheng Chen & Xiaoyuan Wang

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  1. Lianghong Yin
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Correspondence to Xiaoyuan Wang.

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Yin, L., Zhao, J., Chen, C. et al. Enhancing the carbon flux and NADPH supply to increase L-isoleucine production in Corynebacterium glutamicum . Biotechnol Bioproc E 19, 132–142 (2014). https://doi.org/10.1007/s12257-013-0416-z

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  • DOI: https://doi.org/10.1007/s12257-013-0416-z

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