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Engineering Clostridium acetobutylicum with a histidine kinase knockout for enhanced n-butanol tolerance and production

  • Bioenergy and biofuels
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Abstract

Clostridium acetobutylicum JB200, a mutant strain of C. acetobutylicum ATCC 55025 obtained through strain evolution in a fibrous bed bioreactor, had high butanol tolerance and produced up to ~21 g/L butanol from glucose in batch fermentation, an improvement of ~67 % over the parental strain (~12.6 g/L). Comparative genomic analysis revealed a single-base deletion in the cac3319 gene leading to C-terminal truncation in its encoding histidine kinase (HK) in JB200. To study the effects of cac3319 mutation on cell growth and fermentation, the cac3319 gene in ATCC 55025 was disrupted using the ClosTron group II intron-based gene inactivation system. Compared to ATCC 55025, the cac3319 HK knockout mutant, HKKO, produced 44.4 % more butanol (18.2 ± 1.3 vs. 12.6 ± 0.2 g/L) with a 90 % higher productivity (0.38 ± 0.03 vs. 0.20 ± 0.02 g/L h) due to increased butanol tolerance, confirming, for the first time, that cac3319 plays an important role in regulating solvent production and tolerance in C. acetobutylicum. This work also provides a novel metabolic engineering strategy for generating high-butanol-tolerant and high-butanol-producing strains for industrial applications.

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Acknowledgments

This work was supported in part by the National Science Foundation STTR program (Grant No. IIP-1026648). We would like to thank Prof. N.P. Minton and Dr. J.T. Heap (University of Nottingham, UK) for providing the plasmid pAN2 and Hopen Yang for copyediting the manuscript.

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Authors and Affiliations

  1. William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 W. 19th Ave, Columbus, OH, 43210, USA

    Mengmeng Xu, Jingbo Zhao, Le Yu & Shang-Tian Yang

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

    I-Ching Tang

  3. Department of Life Science and Biotechnology, Dalian University of Technology, Dalian, 116024, China

    Chuang Xue

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  1. Mengmeng Xu
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  2. Jingbo Zhao
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  3. Le Yu
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Correspondence to Shang-Tian Yang.

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Xu, M., Zhao, J., Yu, L. et al. Engineering Clostridium acetobutylicum with a histidine kinase knockout for enhanced n-butanol tolerance and production. Appl Microbiol Biotechnol 99, 1011–1022 (2015). https://doi.org/10.1007/s00253-014-6249-7

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