Journal of Industrial Microbiology & Biotechnology

, Volume 40, Issue 9, pp 1057–1066

In silico aided metabolic engineering of Klebsiella oxytoca and fermentation optimization for enhanced 2,3-butanediol production


  • Jong Myoung Park
    • Research and Development CenterGS Caltex Corporation
    • Research and Development CenterGS Caltex Corporation
  • Hee Jong Lee
    • Research and Development CenterGS Caltex Corporation
  • Doyoung Seung
    • Research and Development CenterGS Caltex Corporation
Metabolic Engineering and Synthetic Biology

DOI: 10.1007/s10295-013-1298-y

Cite this article as:
Park, J.M., Song, H., Lee, H.J. et al. J Ind Microbiol Biotechnol (2013) 40: 1057. doi:10.1007/s10295-013-1298-y


Klebsiella oxytoca naturally produces a large amount of 2,3-butanediol (2,3-BD), a promising bulk chemical with wide industrial applications, along with various byproducts. In this study, the in silico gene knockout simulation of K. oxytoca was carried out for 2,3-BD overproduction by inhibiting the formation of byproducts. The knockouts of ldhA and pflB genes were targeted with the criteria of maximization of 2,3-BD production and minimization of byproducts formation. The constructed K. oxytoca ΔldhA ΔpflB strain showed higher 2,3-BD yields and higher final concentrations than those obtained from the wild-type and ΔldhA strains. However, the simultaneous deletion of both genes caused about a 50 % reduction in 2,3-BD productivity compared with K. oxytoca ΔldhA strain. Based on previous studies and in silico investigation that the agitation speed during 2,3-BD fermentation strongly affected cell growth and 2,3-BD synthesis, the effect of agitation speed on 2,3-BD production was investigated from 150 to 450 rpm in 5-L bioreactors containing 3-L culture media. The highest 2,3-BD productivity (2.7 g/L/h) was obtained at 450 rpm in batch fermentation. Considering the inhibition of acetoin for 2,3-BD production, fed-batch fermentations were performed using K. oxytoca ΔldhA ΔpflB strain to enhance 2,3-BD production. Altering the agitation speed from 450 to 350 rpm at nearly 10 g/L of acetoin during the fed-batch fermentation allowed for the production of 113 g/L 2,3-BD, with a yield of 0.45 g/g, and for the production of 2.1 g/L/h of 2,3-BD.


2,3-ButanediolKlebsiella oxytoca In silico aided metabolic engineering Fermentation optimization

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© Society for Industrial Microbiology and Biotechnology 2013