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Metabolic engineering of Clostridium tyrobutyricum for n-butanol production: effects of CoA transferase

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Abstract

The overexpression of CoA transferase (ctfAB), which catalyzes the reaction: acetate/butyrate + acetoacetyl-CoA → acetyl/butyryl-CoA + acetoacetate, was studied for its effects on acid reassimilation and butanol biosynthesis in Clostridium tyrobutyricum (Δack, adhE2). The plasmid pMTL007 was used to co-express adhE2 and ctfAB from Clostridium acetobutylicum ATCC 824. In addition, the sol operon containing ctfAB, adc (acetoacetate decarboxylase), and ald (aldehyde dehydrogenase) was also cloned from Clostridium beijerinckii NCIMB 8052 and expressed in C. tyrobutyricum (Δack, adhE2). Mutants expressing these genes were evaluated for their ability to produce butanol from glucose in batch fermentations at pH 5.0 and 6.0. Compared to C. tyrobutyricum (Δack, adhE2) without expressing ctfAB, all mutants with ctfAB overexpression produced more butanol, with butanol yield increased to 0.22 − 0.26 g/g (vs. 0.10 − 0.13 g/g) and productivity to 0.35 g/l h (vs. 0.13 g/l h) because of the reduced acetate and butyrate production. The expression of ctfAB also resulted in acetone production from acetoacetate through a non-enzymatic decarboxylation.

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Acknowledgments

This work was supported in part by the National Science Foundation STTR program (IIP-1026648). We are grateful to Prof. N. P. Minton, University of Nottingham, UK for providing the donor E. coli CA434 and plasmid pMTL007 used in this study.

Compliance with ethical standards

This research does not involve human participants or animals.

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The authors declare no conflict of interests.

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Authors and Affiliations

  1. William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 151 West Woodruff Ave., Columbus, OH, 43210, USA

    Le Yu, Jingbo Zhao, Mengmeng Xu, Jie Dong, Saju Varghese, Mingrui Yu & Shang-Tian Yang

  2. Bioprocessing Innovative Company, 4734 Bridle Path Ct., Dublin, OH, 43017, USA

    Mingrui Yu & I-Ching Tang

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  1. Le Yu
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Yu, L., Zhao, J., Xu, M. et al. Metabolic engineering of Clostridium tyrobutyricum for n-butanol production: effects of CoA transferase. Appl Microbiol Biotechnol 99, 4917–4930 (2015). https://doi.org/10.1007/s00253-015-6566-5

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