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Enhanced butanol production by increasing NADH and ATP levels in Clostridium beijerinckii NCIMB 8052 by insertional inactivation of Cbei_4110

  • Applied genetics and molecular biotechnology
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Abstract

Clostridium beijerinckii is identified as a promising Clostridium strain for industrialization of acetone and butanol (AB) fermentation. It has been reported that high reducing power levels are associated with high butanol yield. In this study, we regulated reducing power by blocking NAD(P)H consumption in C. beijerinckii NCIMB 8052. Gene Cbei_4110, encoding NADH-quinone oxidoreductase (nuoG), is a subunit of the electron transport chain complex I. After inactivation of gene Cbei_4110, the generated mutant strain exhibited a remarkable increase in glucose utilization ratio and enhanced butanol production to 9.5 g/L in P2 medium containing 30 g/L of glucose. NAD(P)H and ATP levels were also increased by one to two times and three to five times, respectively. Furthermore, a comparative transcriptome analysis was carried out in order to determine the mechanism involved in the enhanced activity of the Cbei_4110-inactivated mutant strain. This strategy may be extended for making industrial bio-butanol more economically attractive.

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Acknowledgments

The authors would like to thank Prof. Nigel P. Minton and Michelle L. Kelly from the University of Nottingham, UK, for kindly providing the ClosTron plasmids and Prof. Sheng Yang from the Shanghai Institutes for Biological Sciences for kindly providing the pWJ1 plasmid.

This work was supported by the National Basic Research Program of China (973 Program, 2013CB733602), the restructured institutions innovation capacity of special funds of Ministry of Science and Technology of China (Grant No. 2014EG111227), the National Natural Science Foundation of China (Grant No. 21306032), the National High-Tech Research and Development Program of China (863) (Grant No. 2012AA021200), the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD), Postdoctoral Science Foundation of Jiangsu Province (Grant No. 1302107C), the Major Research Plan of the National Natural Science Foundation of China (21390204), Program for Changjiang Scholars and Innovative Research Team in University (Grant No.: IRT_14R28), and Research Foundation of Guangzhou Sugarcane Industry Research Institute (Grant No. A201302).

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Authors and Affiliations

  1. State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nan**g Tech University, No. 30, Puzhu South Road, Nan**g, 211816, China

    Jun Liu, Ting Guo, Dong Wang, **aoning Shen, Dong Liu, Huanqing Niu & Hanjie Ying

  2. National Engineering Technique Research Center for Biotechnology, Nan**g, 211816, People’s Republic of China

    Jun Liu, Ting Guo, Dong Wang, **aoning Shen, Dong Liu, Huanqing Niu & Hanjie Ying

  3. Guangzhou Sugarcane Industry Research Institute, Guangdong Key Laboratory of Sugarcane Improvement and Biorefinery, Guangdong Engineering Research & Development Center for Comprehensive Utilization of Plant Fiber, Guangzhou Key Laboratory for Comprehensive Utilization of Plant Fiber, No. 10, Shiliu gang Road, Guangzhou, 510316, China

    Ting Guo & Lei Liang

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  1. Jun Liu
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Jun Liu, Ting Guo and Dong Wang contributed equally to this work.

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Liu, J., Guo, T., Wang, D. et al. Enhanced butanol production by increasing NADH and ATP levels in Clostridium beijerinckii NCIMB 8052 by insertional inactivation of Cbei_4110. Appl Microbiol Biotechnol 100, 4985–4996 (2016). https://doi.org/10.1007/s00253-016-7299-9

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