Applied Microbiology and Biotechnology

, Volume 87, Issue 3, pp 943–949

Enhanced production of 2,3-butanediol from glycerol by forced pH fluctuations

Biotechnological Products and Process Engineering


The glycerol fermentation by Klebsiella pneumoniae occurs by receiving more than five liquid products—organic acids, diols, and ethanol. Aiming to direct the glycerol conversion towards predominant production of 2,3-butanediol (2,3-BD), the main influencing parameters (the aeration and the pH) were investigated during fed-batch processes. The regime of intensive aeration (2.2 vvm air supply) was evaluated as most favorable for 2,3-BD synthesis and ensured the decrease of all other metabolites. Thus, without pH control, 52.5 g/l 2,3-BD were produced, as the carbon conversion of glycerol into 2,3-BD reached 60.6%. Additional enhancement in 2,3-BD production (by significant increase of glycerol utilization) was achieved by the development of a new method of “forced pH fluctuations”. It was realized by consecutive raisings of pH using definite ΔpH value, at exact time intervals, allowing multiple variations. Thus, the optimal conditions for maximal glycerol consumption were defined, and 70 g/l 2,3-BD were produced, which is the highest amount obtained from glycerol as a sole carbon source until now. The forced pH fluctuations emphasized pH as a governing factor in microbial conversion processes.


2,3-Butanediol Glycerol Klebsiella pneumoniae pH fluctuation 


  1. Biebl H, Menzel K, Zeng AP, Deckwer WD (1999) Microbial production of 1, 3-propanediol. Appl Microbiol Biotechnol 52:289–297CrossRefGoogle Scholar
  2. Cheng KK, Liu DH, Sun Y, Liu WB (2004) 1, 3-Propanediol production by Klebsiella pneumoniae under different aeration strategies. Biotechnol Lett 26:911–915CrossRefGoogle Scholar
  3. Da Silva GP, Mack M, Contiero J (2009) Glycerol: a promising and abundant carbon source for industrial microbiology. Biotechnol Adv 27:30–39CrossRefGoogle Scholar
  4. Ji XJ, Huang H, Li S, Du J, Lian M (2008) Enhanced 2, 3-butanediol production by altering the mixed acid fermentation pathway in Klebsiella oxytoca. Biotechnol Lett 30:731–734CrossRefGoogle Scholar
  5. Ji XJ, Huang H, Du J, Zhu JG, Ren LJ, Li S, Nie ZK (2009a) Development of industrial medium for economical 2, 3-butanediol production through co-fermentation of glucose and xylose by Klebsiella oxytoca. Biores Technol 100:5214–5218CrossRefGoogle Scholar
  6. Ji XJ, Huang H, Zhu JG, Ren LJ, Nie ZK, Du J, Li S (2009b) Engineering Klebsiella oxytoca for efficient 2, 3-butanediol production through insertional inactivation of acetaldehyde dehydrogenase gene. Appl Microbiol Biotechnol. doi:10.1007/s00253-009-2222-2 Google Scholar
  7. Ji XJ, Huang H, Zhu JG, Hu N, Li S (2009c) Efficient 1, 3-propanediol production by fed-batch culture of Klebsiella pneumoniae: the role of pH fluctuations. Appl Biochem Biotechnol. doi:10.1007/s12010-008-8492-9 Google Scholar
  8. Ma C, Wang A, Qin J, Li L, Ai X, Jiang T, Tang H, Xu P (2009) Enhanced 2,3-butanediol production by Klebsiella pneumoniae SDM. Appl Microbiol Biotechnol 82:49–57Google Scholar
  9. Petrova P, Petrov K, Beschkov V (2009) Production of 1, 3-propanediol from glycerol by newly isolated strains of Klebsiella pneumoniae. Compt Rend Acad Bulg Sci 62:233–242Google Scholar
  10. Petrov K, Petrova P (2009) High production of 2, 3-butanediol from glycerol by Klebsiella pneumoniae G31. Appl Microbiol Biotechnol 84:659–665CrossRefGoogle Scholar
  11. Qin J, Xiao Z, Ma C, Xie N, Liu P, Xu P (2006) Production of 2, 3-butanediol by Klebsiella pneumoniae using glucose and ammonium phosphate. Chi J Chem Eng 14:132–136CrossRefGoogle Scholar
  12. Sun LH, Wang XD, Dai JY, Xiu ZL (2009) Microbial production of 2, 3-butanediol from Jerusalem artichoke tubers by Klebsiella pneumoniae. Appl Microbiol Biotechnol 82:847–852CrossRefGoogle Scholar
  13. Syu MJ (2001) Biological production of 2, 3-butanediol. Appl Microbiol Biotechnol 55:10–18CrossRefGoogle Scholar
  14. Wu KJ, Saratale GD, Lo YC, Chen WM, Tseng ZJ, Chang MC, Tsai BC, Su A, Chang JS (2008) Simultaneous production of 2, 3-butanediol, ethanol and hydrogen with a Klebsiella sp. strain isolated from sewage sludge. Bioresour Technol 99:7966–7970CrossRefGoogle Scholar
  15. Xiu ZL, Zeng AP (2008) Present state and perspective of downstream processing of biologically produced 1, 3-propanediol and 2, 3-butanediol. Appl Microbiol Biotechnol 78:917–926CrossRefGoogle Scholar
  16. Yang G, Tian J, Li J (2007) Fermentation of 1, 3-propanediol by a lactate deficient mutant of Klebsiella oxytoca under microaerobic conditions. Appl Microbiol Biotechnol 73:1017–1024CrossRefGoogle Scholar
  17. Zeng AP, Biebl H (2002) Bulk chemicals from biotechnology: the case of 1, 3-propanediol production and the new trends. Adv Biochem Eng Biotechnol 74:239–259Google Scholar
  18. Zeng AP, Biebl H, Deckwer WD (1990) Effect of pH and acetic acid on growth and 2, 3-butanediol production of Enterobacter aerogenes in continuous culture. Appl Microbiol Biotechnol 33:485–489Google Scholar
  19. Zheng ZM, Xu YZ, Liu HJ, Guo NN, Cai ZZ, Liu DH (2008) Physiologic mechanisms of sequential products synthesis in 1, 3-propanediol fed-batch fermentation by Klebsiella pneumoniae. Biotechnol Bioeng 100:923–932CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  1. 1.Institute of Chemical EngineeringBulgarian Academy of SciencesSofiaBulgaria
  2. 2.Institute of MicrobiologyBulgarian Academy of SciencesSofiaBulgaria

Personalised recommendations