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Reconstructing the clostridial n-butanol metabolic pathway in Lactobacillus brevis

  • Applied Genetics and Molecular Biotechnology
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Abstract

A Lactobacillus brevis strain with the ability to synthesize butanol from glucose was constructed by metabolic engineering. The genes crt, bcd, etfB, etfA, and hbd, composing the bcs-operon, and the thl gene encode the enzymes of the lower part of the clostridial butanol pathway (crotonase, butyryl-CoA-dehydrogenase, two subunits of the electron transfer flavoprotein, 3-hydroxybutyryl-CoA dehydrogenase, and thiolase) of Clostridium acetobutylicum. They were cloned into the Gram-positive/Gram-negative shuttle plasmid vector pHYc. The two resulting plasmids pHYc-thl-bcs and pHYc-bcs (respectively, with and without the clostridial thl gene) were transferred to Escherichia coli and L. brevis. The recombinant L. brevis strains were able to synthesize up to 300 mg l−1 or 4.1 mM of butanol on a glucose-containing medium. A L. brevis strain carrying the clostridial bcs-operon has the ability to synthesize butanol with participation of its own thiolase, aldehyde dehydrogenase, and alcohol dehydrogenase. The particular role of the enzymes involved in butanol production and the suitability of L. brevis as an n-butanol producer are discussed.

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Acknowledgments

This study was supported by the Bioprocess Group, Moscow. Dr. Iijima kindly provided the shuttle-vector pHYc utilized for expression of clostridial genes as a gift. We are most grateful to Dr. Rustem Shakulov for his invaluable advice and Olesya Shashkina for technical assistance.

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Authors and Affiliations

  1. State Research Institute of Genetics and Selection of Industrial Microorganisms, 1st Dorojniy pr. 1, 117545, Moscow, Russian Federation

    Oksana V. Berezina, Natalia V. Zakharova & Sergey V. Yarotsky

  2. Department of Microbiology, Technische Universität München, Emil-Ramann-Str. 4, 85350, Freising, Germany

    Agnieszka Brandt, Wolfgang H. Schwarz & Vladimir V. Zverlov

  3. Institute of Molecular Genetics, Russian Academy of Science, Kurchatov Sq. 2, 123182, Moscow, Russian Federation

    Vladimir V. Zverlov

Authors
  1. Oksana V. Berezina
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  2. Natalia V. Zakharova
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  3. Agnieszka Brandt
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  4. Sergey V. Yarotsky
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  5. Wolfgang H. Schwarz
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  6. Vladimir V. Zverlov
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Correspondence to Oksana V. Berezina or Natalia V. Zakharova.

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Oksana V. Berezina and Natalia V. Zakharova equally contributed to this work.

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Berezina, O.V., Zakharova, N.V., Brandt, A. et al. Reconstructing the clostridial n-butanol metabolic pathway in Lactobacillus brevis . Appl Microbiol Biotechnol 87, 635–646 (2010). https://doi.org/10.1007/s00253-010-2480-z

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  • DOI: https://doi.org/10.1007/s00253-010-2480-z

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