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Enhanced production of 2,3-butanediol by engineered Bacillus subtilis

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Abstract

Production of 2,3-butanediol by Bacillus subtilis takes place in late-log or stationary phase, depending on the expression of bdhA gene encoding acetoin reductase, which converts acetoin to 2,3-butanediol. The present work focuses on the development of a strain of B. subtilis for enhanced production of 2,3-butanediol in early log phase of growth cycle. For this, the bdhA gene was expressed under the control of P alsSD promoter of AlsSD operon for acetoin fermentation which served the substrate for 2,3-butanediol production. Addition of acetic acid in the medium induced the production of 2,3-butanediol by 2-fold. Two-step aerobic–anaerobic fermentation further enhanced 2,3-butanediol production by 4-fold in comparison to the control parental strain. Thus, addition of acetic acid and low dissolved oxygen in the medium are involved in activation of bdhA gene expression from P alsSD promoter in early log phase. Under the conditions tested in this work, the maximum production of 2,3-butanediol, 2.1 g/l from 10 g/l glucose, was obtained at 24 h. Furthermore, under the optimized microaerophilic condition, the production of 2,3-butanediol improved up to 6.1 g/l and overall productivity increased by 6.7-fold to 0.4 g/l h in the engineered strain compared to that in the parental control.

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Acknowledgements

This work was supported by a financial grant from the Special Coordination Funds for Promoting Science and Technology and the Creation of Innovation Centers for Advanced Interdisciplinary Research Areas (Innovative Bioproduction Kobe) Programs and in part by Advanced Low Carbon Technology Research and Development Program of the Ministry of Education, Science and Sports and Culture of Japan. We thank Dr. Pulla Kaothien-Nakayama for critical reading of the paper.

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Authors and Affiliations

  1. Organization of Advance Science and Technology, Kobe University, Kobe, 657-8501, Japan

    Ranjita Biswas, Hideki Nakayama & Takashi Kondo

  2. Department of Agrobioscience, Graduate School of Agricultural Science, Kobe University, Kobe, 657-8501, Japan

    Masaru Yamaoka & Ken-ichi Yoshida

  3. Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi, New Delhi, 110016, India

    Virendra S. Bisaria

  4. Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan

    Akihiko Kondo

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  1. Ranjita Biswas
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  2. Masaru Yamaoka
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  3. Hideki Nakayama
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  4. Takashi Kondo
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  5. Ken-ichi Yoshida
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  6. Virendra S. Bisaria
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  7. Akihiko Kondo
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Correspondence to Akihiko Kondo.

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Biswas, R., Yamaoka, M., Nakayama, H. et al. Enhanced production of 2,3-butanediol by engineered Bacillus subtilis . Appl Microbiol Biotechnol 94, 651–658 (2012). https://doi.org/10.1007/s00253-011-3774-5

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  • DOI: https://doi.org/10.1007/s00253-011-3774-5

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