Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Continuous production of isopropanol and butanol using Clostridium beijerinckii DSM 6423


Clostridium beijerinckii DSM 6423 was studied using different continuous production methods to give maximum and stable production of isopropanol and n-butanol. In a single-stage continuous culture, when wood pulp was added as a cell holding material, we could increase the solvent productivity from 0.47 to 5.52 g L−1 h−1 with the yield of 54% from glucose. The overall solvent concentration of 7.51 g L−1 (39.4% isopropanol and 60.6% n-butanol) with the maximum solvent productivity of 0.84 g L−1 h−1 was obtained with two-stage continuous culture. We were able to run the process for more than 48 overall retention times without losing the ability to produce solvents.

This is a preview of subscription content, log in to check access.

We’re sorry, something doesn't seem to be working properly.

Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.

Fig. 1
Fig. 2
Fig. 3
Fig. 4


  1. Ahmed I, Ross RA, Mathur VK, Chesbro WR (1988) Growth rate dependence of solventogenesis and solvents produced by Clostridium beijerinckii. Appl Microbiol Biotechnol 28:182–187

  2. Bahl H, Andersch W, Gottschalk G (1982) Continuous production of acetone and butanol by Clostridium acetobutylicum in a two-stage phosphate limited chemostat. Appl Microbiol Biotechnol 15(4):201–205

  3. Cascone R (2008) Biobutanol—a replacement for bioethanol? Chem Eng Prog 104:S4–S9

  4. Chen JS, Hiu SF (1986) Acetone–butanol–isopropanol production by Clostridium beijerinckii (Synonym, Clostridium butylicum). Biotechnol Lett 8(5):371–376

  5. Ennis BM, Maddox IS (1989) Production of solvents (ABE fermentation) from whey permeate by continuous fermentation in a membrane bioreactor. Bioprocess Eng 4:27–34

  6. Ezeji T, Blaschek HP (2008) Fermentation of dried distillers' grains and solubles (DDGS) hydrolysates to solvents and value-added products by solventogenic clostridia. Bioresour Technol 99(12):5232–5242

  7. Forsberg CW (1987) Production of 1, 3-propanediol from glycerol by Clostridium acetobutylicum and other Clostridium species. Appl Environ Microbiol 53:639–643

  8. Gapes JR, Nimcevic D, Friedl A (1996) Long-term continuous cultivation of Clostridium beijerinckii in a two-stage chemostat with on-line solvent removal. Appl Environ Microbiol 62(9):3210–3219

  9. George HA, Chen JS (1983) Acidic conditions are not obligatory for onset of butanol formation by Clostridium beijerinckii (Synonym, C. butylicum). Appl Environ Microbiol 46(2):321–327

  10. George HA, Johnson JL, Moore WEC, Holdeman LV, Chen JS (1983) Acetone, isopropanol, and butanol production by Clostridium beijerinckii (syn. Clostridium butylicum) and Clostridium aurantibutyricum. Appl Environ Microbiol 45(3):1160–1163

  11. Ghosh S, Klass DL (1978) Two-phase anaerobic digestion. Process Biochem 13(4):15–24

  12. Godin C, Engasser JM (1989) Acid in the first stage is a determinant factor for the solvent production in the two-stage continuous fermentation of Clostridium acetobutylicum. Biotechnol Lett 11:903–906

  13. Green EM (2011) Fermentative production of butanol—the industrial perspective. Curr Opin Biotechnol 22:1–7

  14. Harris LM, Desai RP, Welker NE, Papoutsakis ET (2000) Characterization of recombinant strains of the Clostridium acetobutylicum butyrate kinase inactivation mutant: need for new phenomenological models for solventogenesis and butanol inhibition? Biotechnol Bioeng 67:1–11

  15. Harris LM, Blank L, Desai RP, Welker NE, Papoutsakis ET (2001) Fermentation characterization and flux analysis of recombinant strains of Clostridium acerobutylicum with an inactivated solR gene. J Ind Microbiol Biotechnol 27:322–328

  16. Huang WC, Ramey D, Yang ST (2004) Continuous production of butanol by Clostridium acetobutylicum immobilized in a fibrous bed bioreactor. Appl Biochem Biotechnol 115(1):887–898

  17. Johnson JL, Toth J, Santiwatanakul S, Chen JS (1997) Cultures of “Clostridium acetobutylicum” from various collections comprise Clostridium acetobutylicum, Clostridium beijerinckii, and two other distinct types based on DNA–DNA reassociation. Int J Syst Bacteriol 47:420–424

  18. Keis S, Bennett CF, Ward VK, Jones DT (1995) Taxonomy and phylogeny of industrial solvent-producing clostridia. Int J Syst Bacteriol 45:693–705

  19. Krouwel PG, van der Laan WFM, Kossen NWF (1980) Continuous production of n-butanol and isopropanol by immobilized, growing Clostridium butylicum cells. Biotechnol Lett 2(5):253–258

  20. Lee SM, Cho MO, Park CH, Chung YC, Kim JH, Sang BI, Um Y (2008) Continuous butanol production using suspended and immobilized Clostridium beijerinckii NCIMB 8052 with supplementary butyrate. Energy Fuels 22(5):3459–3464

  21. Li SY, Srivastava R, Suib SL, Li Y, Parnas RS (2011) Performance of batch, fed-batch, and continuous A–B–E fermentation with pH-control. Biores Technol 102(5):4241–4250

  22. Lütke-Eversloh T, Bahl H (2011) Metabolic engineering of Clostridium acetobutylicum: recent advances to improve butanol production. Curr Opin Biotechnol. doi:

  23. Maddox IS (1989) The acetone–butanol–ethanol fermentation: recent progress in technology. Biotechnol Genet Eng Rev 7:189–220

  24. Mariano AP, Costa CBB, De Angelis DDF, Maugeri Filho F, Atala DIP, Wolf Maciel MR, Maciel Filho R (2010) Optimisation of a fermentation process for butanol production by particle swarm optimisation (PSO). J Chem Technol Biotechnol 85(7):934–949

  25. Mulchaldani A, Volesky B (1994) Production of acetone–butanol–ethanol by Clostridium acetobutylicum using a spin filter perfusion bioreactor. J Biotechnol 34:51–60

  26. Mutschlechner O, Swoboda H, Gapes JR (2000) Continuous two-stage ABE-fermentation using Clostridium beijerinckii NRRL B592 operating with a growth rate in the first stage vessel close to its maximal value. J Mol Microbiol Biotechnol 2(1):101–105

  27. Nair RV, Bennett GN, Papoutsakis ET (1994) Molecular characterization of an aldehyde/alcohol dehydrogenase gene from Clostridium acetobutylicum ATCC 824. J Bacteriol 176:871–885

  28. Nielsen DR, Prather KJ (2009) In situ product recovery of n-butanol using polymeric resins. Biotechnol Bioeng 102(3):811–821

  29. Palsson BO, Fathi-Afshar S, Rudd DF, Lightfoot EN (1981) Biomass as a source of chemical feedstocks. Science 213:513–517

  30. Papanikolaou S, Fakas S, Fick M, Chevalot I, Galiotou-Panayotou M, Komaitis M, Marc I, Aggelis G (2008) Biotechnological valorisation of raw glycerol discharged after bio-diesel (fatty acid methyl esters) manufacturing process: production of 1,3-propanediol, citric acid and single cell oil. Biomass Bioenerg 32(1):60–71

  31. Qureshi N, Maddox IS (1995) Continuous production of acetone–butanol–ethanol using immobilized cells of Clostridium acetobutylicum and integration with product removal by liquid–liquid extraction. J Ferment Bioeng 80(2):185–189

  32. Qureshi N, Schripsema J, Lienhardt J, Blaschek HP (2000) Continuous solvent production by Clostridium beijerinckii BA101 immobilized by adsorption onto brick. World J Microbiol Biotechnol 16(4):377–382

  33. Qureshi N, Lai LL, Blaschek HP (2004) Scale-up of a high productivity continuous biofilm reactor to produce butanol by adsorbed cells of Clostridium beijerinckii. Trans IChemE Part C 82(C2):164–173

  34. Qureshi N, Ezeji TC, Ebener J, Dien BS, Cotta MA, Blaschek HP (2008) Butanol production by Clostridium beijerinckii. Part I: use of acid and enzyme hydrolyzed corn fiber. Bioresour Technol 99(13):5915–5922

  35. Sun Z, Liu S (2010) Production of n-butanol from concentrated sugar maple hemicellulosic hydrolysate by Clostridia acetobutylicum ATCC824. Biom Bioener. doi:

  36. Tashiro Y, Takeda K, Kobayashi G, Sonomoto K (2005) High production of acetone–butanol–ethanol with high cell density culture by cell-recycling and bleeding. J Biotechnol 120(2):197–206

  37. Tomas CA, Welker NE, Papoutsakis ET (2003) Overexpression of groESL in Clostridium acetobutylicum results in increased solvent production and tolerance, prolonged metabolism, and changes in the cell's transcriptional program. Appl Environ Microbiol 69:4951–4965

  38. Wilkinson SR, Young M, Goodacre R, Morris JG, Farrow JAE, Collins MD (1995) Phenotypic and genotypic differences between certain strains of Clostridium acetobutylicum. FEMS Microbiol Lett 125:199–204

  39. Yan RT, Zhu CX, Golemboski C, Chen JS (1988) Expression of solvent-forming enzymes and onset of solvent production in batch cultures of Clostridium beijerinckii (“Clostridium butylicum”). Appl Environ Microbiol 54:642–648

  40. Zhang Y, Ma Y, Yang F, Zhang C (2009) Continuous acetone–butanol–ethanol production by corn stalk immobilized cells. J Ind Microbiol Biotechnol 36:1117–1121

Download references

Author information

Correspondence to Tom Granström.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Survase, S.A., Jurgens, G., van Heiningen, A. et al. Continuous production of isopropanol and butanol using Clostridium beijerinckii DSM 6423. Appl Microbiol Biotechnol 91, 1305–1313 (2011).

Download citation


  • Isopropanol
  • n-butanol
  • Chemostat
  • Clostridium beijerinckii