Applied Microbiology and Biotechnology

, Volume 93, Issue 3, pp 1057–1063 | Cite as

High-level production of 1,3-propanediol from crude glycerol by Clostridium butyricum AKR102a

  • Erik Wilkens
  • Anne Katrin Ringel
  • Diana Hortig
  • Thomas WillkeEmail author
  • Klaus-Dieter Vorlop
Biotechnological products and process engineering


The aim of this study was to optimize a biotechnological process for the production of 1,3-propanediol (1,3-PD) based on low-quality crude glycerol derived from biodiesel production. Clostridium butyricum AKR102a was used in fed-batch fermentations in 1-L and 200-L scale. The newly discovered strain is characterized by rapid growth, high product tolerance, and the ability to use crude glycerol at the lowest purity directly gained from a biodiesel plant side stream. Using pure glycerol, the strain AKR102 reached 93.7 g/L 1,3-PD with an overall productivity of 3.3 g/(L*h). With crude glycerol under the same conditions, 76.2 g/L 1,3-PD was produced with a productivity of 2.3 g/(L*h). These are among the best results published so far for natural producers. The scale up to 200 L was possible. Due to the simpler process design, only 61.5 g/L 1,3-PD could be reached with a productivity of 2.1 g/(L*h).


Fermentation Crude glycerol 1,3-Propanediol Clostridium butyricum 



This project was funded by the German Federal Ministry of Education and Research (BMBF) (IG-Biotech 0315026E). I want to thank PD Dr. Udo Rau and Wolfgang Graßl from the Technische Universität Braunschweig for the kind technical provision of the large-scale fermenter and the assistance during the 200-L fermentation.


  1. Barbirato F, Himmi EH, Conte T, Bories A (1998) 1,3-Propanediol production by fermentation: an interesting way to valorize glycerin from the ester and ethanol industries. Ind Crop Prod 7:281–289CrossRefGoogle Scholar
  2. Belcher LA, Muska CF, DeSalvo JW (2010) Evaluating 1,3-propanediol for potential skin effects. Cosmet Toilet 125:81–86Google Scholar
  3. Biebl H, Marten S, Hippe H, Deckwer WD (1992) Glycerol conversion to 1,3-propanediol by newly isolated clostridia. Appl Microbiol Biotechnol 36:592–597CrossRefGoogle Scholar
  4. Bock R (2004) Biokonversion von Glycerin zu 1,3-Propandiol mit freien und immobilisierten Mikroorganismen. Technische Universität Braunschweig, DissertationGoogle Scholar
  5. Boenigk R, Bowien S, Gottschalk G (1993) Fermentation of glycerol to 1,3-propanediol in continuous cultures of Citrobacter freundii. Appl Microbiol Biotechnol 38:453–457CrossRefGoogle Scholar
  6. Chatzifragkou A, Dietz D, Komaitis M, Zeng AP, Papanikolaou S (2010) Effect of biodiesel-derived waste glycerol impurities on biomass and 1,3-propanediol production of Clostridium butyricum VPI 1718. Biotechnol Bioeng 107:76–84CrossRefGoogle Scholar
  7. Chatzifragkou A, Papanikolaou S, Dietz D, Doulgeraki AI, Nychas GJE, Zeng AP (2011) Production of 1,3-propanediol by Clostridium butyricum growing on biodiesel-derived crude glycerol through a non-sterilized fermentation process. Appl Microbiol Biotechnol 91:101–112CrossRefGoogle Scholar
  8. 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
  9. Cheng KK, Zhang JA, Liu DH, Sun Y, Liu HJ, Yang MD, Xu JM (2007) Pilot-scale production of 1,3-propanediol using Klebsiella pneumoniae. Process Biochem 42:740–744CrossRefGoogle Scholar
  10. Colin T, Bories A, Lavigne C, Moulin G (2001) Effects of acetate and butyrate during glycerol fermentation by Clostridium butyricum. Curr Microbiol 43:238–243CrossRefGoogle Scholar
  11. Dabrock B, Bahl H, Gottschalk G (1992) Parameters affecting solvent production by Clostridium pasteurianum. Appl Environ Microbiol 58:1233–1239Google Scholar
  12. Eaton RE, Boon WH, Smith CJ (2004) Chemical base for engine coolant/antifreeze with improved thermal stability properties. Patent Num US 6818146 B2 Shell Oil CompanyGoogle Scholar
  13. González-Pajuelo M, Andrade JC, Vasconcelos I (2004) Production of 1,3-propanediol by Clostridium butyricum VPI 3266 using a synthetic medium and raw glycerol. J Ind Microbiol Biotechnol 31:442–446CrossRefGoogle Scholar
  14. Günzel B, Yonsel S, Deckwer WD (1991) Fermentative production of 1,3-propanediol from glycerol by Clostridium butyricum up to a scale of 2 m³. Appl Microbiol Biotechnol 36:289–294CrossRefGoogle Scholar
  15. Hartlep M, Zeng AP (2002) Fedbatch-Verfahren für die mikrobielle Herstellung von 1,3-Propandiol in Klebsielle pneumoniae und Clostridium butyricum. Chem Ing Tech 75:663–664CrossRefGoogle Scholar
  16. Hirschmann S, Baganz K, Koschik I, Vorlop KD (2005) Development of an integrated bioconversion process for the production of 1,3-propanediol from raw glycerol waters. Landbauforsch Volkenrode 55:261–267Google Scholar
  17. Homann T, Tag C, Biebl H, Deckwer WD, Schink B (1990) Fermentation of glycerol to 1,3-propanediol by Klebsiella and Citrobacter strains. Appl Microbiol Biotechnol 33:121–126CrossRefGoogle Scholar
  18. Jun SA, Moon C, Kang CH, Kong SW, Sang BI, Um Y (2010) Microbial fed-batch production of 1,3-propanediol using raw glycerol with suspended and immobilized Klebsiella pneumoniae. Appl Biochem Biotechnol 161:491–501CrossRefGoogle Scholar
  19. Kraus GA (2008) Synthetic methods for the preparation of 1,3-propanediol. Clean Soil Air Water 36:648–651CrossRefGoogle Scholar
  20. Liu HJ, Zhang DJ, Xu YH, Mu Y, Sun YQ, Xiu ZL (2007) Microbial production of 1,3-propanediol from glycerol by Klebsiella pneumoniae under micro-aerobic conditions up to a pilot scale. Biotechnol Lett 29:1281–1285CrossRefGoogle Scholar
  21. Liu HJ, Ou XJ, Zhou S, Liu DH (2010) Microbial 1,3-propanediol, its copolymerization with terephthalate, and applications. Microbiology Monographs 14:405–425CrossRefGoogle Scholar
  22. Moon C, Ahn JH, Kim SW, Sang BI, Um Y (2010) Effect of biodiesel-derived raw glycerol on 1,3-propanediol production by different microorganisms. Appl Biochem Biotechnol 161:502–510CrossRefGoogle Scholar
  23. Mu Y, Teng H, Zhang DJ, Wang W, Xiu ZL (2006) Microbial production of 1,3-propanediol by Klebsiella pneumoniae using crude glycerol from biodiesel preparations. Biotechnol Lett 28:1755–1759CrossRefGoogle Scholar
  24. Nakamura CE, Whited GM (2003) Metabolic engineering for the microbial production of 1,3-propanediol. Curr Opin Biotechnol 14:454–459CrossRefGoogle Scholar
  25. Otte B, Grunwaldt E, Mahmoud O, Jennewein S (2009) Genome shuffling in Clostridium diolis DSM 15410 for improved 1,3-propanediol production. Appl Environ Microbiol 75:7610–7616CrossRefGoogle Scholar
  26. Petitdemange E, Durr C, Andaloussi SA, Raval G (1995) Fermentation of raw glycerol to 1,3-propanediol by new strains of Clostridium butyricum. J Ind Microbiol 15:498–502CrossRefGoogle Scholar
  27. Rehman AU, Matsumura M, Nomura N, Sato S (2008) Growth and 1,3-propanediol production on pre-treated sunflower oil bio-diesel raw glycerol using a strict anaerobe-Clostridium butyricum. Curr Res Bacteriol 1:7–16CrossRefGoogle Scholar
  28. Ringel AK, Wilkens E, Hortig D, Willke T, Vorlop KD (2011) An improved screening method for microorganisms able to convert crude glycerol to 1,3-propanediol and to tolerate high product concentrations. Applied Microbiology and Biotechnology doi: 10.1007/s00253-011-3594-7
  29. Schuetz H, Radler F (1984) Anaerobic reduction of glycerol to 1,3-propanediol by Lactobacillus brevis and Lactobacillus buchneri. Syst Appl Microbiol 5:169–178Google Scholar
  30. Tang XM, Tan YS, Zhu H, Zhao K, Shen W (2009) Microbial conversion of glycerol to 1,3-propanediol by an engineered strain of Escherichia coli. Appl Environ Microbiol 75:1628–1634CrossRefGoogle Scholar
  31. Xu YZ, Guo NN, Zheng ZM, Ou XJ, Liu HJ, Liu DH (2009) Metabolism in 1,3-propanediol fed-batch fermentation by a D-lactate deficient mutant of Klebsiella pneumoniae. Biotechnol Bioeng 104:965–972CrossRefGoogle Scholar
  32. Yazdani SS, Gonzalez R (2007) Anaerobic fermentation of glycerol: a path to economic viability for the biofuels industry. Curr Opin Biotechnol 18:213–219CrossRefGoogle Scholar
  33. Zeng AP (1996) Pathway and kinetic analysis of 1,3-propanediol production from glycerol fermentation by Clostridium butyricum. Bioprocess Eng 14:169–175CrossRefGoogle Scholar
  34. Zeng AP, Biebl H (2002) Bulk chemicals from biotechnology: the case of 1,3-propanediol production and new trends. Adv Biochem Eng Biotechnol 74:239–259Google Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Erik Wilkens
    • 1
  • Anne Katrin Ringel
    • 1
  • Diana Hortig
    • 1
  • Thomas Willke
    • 1
    Email author
  • Klaus-Dieter Vorlop
    • 1
  1. 1.Johann Heinrich von Thünen-Institut (vTI)Institute of Agricultural Technology and Biosystems Engineering (AB)BraunschweigGermany

Personalised recommendations