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Effects of different replicons in conjugative plasmids on transformation efficiency, plasmid stability, gene expression and n-butanol biosynthesis in Clostridium tyrobutyricum

  • Bioenergy and biofuels
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Abstract

Clostridium tyrobutyricum ATCC 25755 can produce butyric acid, acetic acid, and hydrogen as the main products from various carbon sources. In this study, C. tyrobutyricum was used as a host to produce n-butanol by expressing adhE2 gene under the control of a native thiolase promoter using four different conjugative plasmids (pMTL82151, 83151, 84151, and 85151) each with a different replicon (pBP1 from C. botulinum NCTC2916, pCB102 from C. butyricum, pCD6 from Clostridium difficile, and pIM13 from Bacillus subtilis). The effects of different replicons on transformation efficiency, plasmid stability, adhE2 expression and aldehyde/alcohol dehydrogenase activities, and butanol production by different mutants of C. tyrobutyricum were investigated. Among the four plasmids and replicons studied, pMTL82151 with pBP1 gave the highest transformation efficiency, plasmid stability, gene expression, and butanol biosynthesis. Butanol production from various substrates, including glucose, xylose, mannose, and mannitol were then investigated with the best mutant strain harboring adhE2 in pMTL82151. A high butanol titer of 20.5 g/L with 0.33 g/g yield and 0.32 g/L h productivity was obtained with mannitol as the substrate in batch fermentation with pH controlled at ~6.0.

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Acknowledgments

This work was supported by the National Science Foundation STTR program (IIP-0810568, IIP-1026648) and the Ohio Department of Development—Third Frontier Advanced Energy Program (Tech 08–036). We would like to thank Prof. Minton of University of Nottingham, UK for providing the donor E. coli CA434 and plasmids pMTL82151-85151.

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Authors and Affiliations

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

    Mingrui Yu, Yinming Du, Wenyan Jiang, Wei-Lun Chang & Shang-Tian Yang

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

    I-Ching Tang

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  1. Mingrui Yu
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  2. Yinming Du
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  3. Wenyan Jiang
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  4. Wei-Lun Chang
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Correspondence to Shang-Tian Yang.

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Yu, M., Du, Y., Jiang, W. et al. Effects of different replicons in conjugative plasmids on transformation efficiency, plasmid stability, gene expression and n-butanol biosynthesis in Clostridium tyrobutyricum . Appl Microbiol Biotechnol 93, 881–889 (2012). https://doi.org/10.1007/s00253-011-3736-y

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  • DOI: https://doi.org/10.1007/s00253-011-3736-y

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