Applied Microbiology and Biotechnology

, Volume 94, Issue 3, pp 651–658 | Cite as

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

  • Ranjita Biswas
  • Masaru Yamaoka
  • Hideki Nakayama
  • Takashi Kondo
  • Ken-ichi Yoshida
  • Virendra S. Bisaria
  • Akihiko Kondo
Biotechnological products and process engineering


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.


2,3-Butanediol Bacillus subtilis Anaerobic fermentation bdhA gene Microaerophilic 



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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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Ranjita Biswas
    • 1
  • Masaru Yamaoka
    • 2
  • Hideki Nakayama
    • 1
  • Takashi Kondo
    • 1
  • Ken-ichi Yoshida
    • 2
  • Virendra S. Bisaria
    • 3
  • Akihiko Kondo
    • 4
  1. 1.Organization of Advance Science and TechnologyKobe UniversityKobeJapan
  2. 2.Department of Agrobioscience, Graduate School of Agricultural ScienceKobe UniversityKobeJapan
  3. 3.Department of Biochemical Engineering and BiotechnologyIndian Institute of Technology DelhiNew DelhiIndia
  4. 4.Department of Chemical Science and Engineering, Graduate School of EngineeringKobe UniversityKobeJapan

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