Applied Microbiology and Biotechnology

, Volume 93, Issue 4, pp 1485–1494 | Cite as

Continuous butanol production with reduced byproducts formation from glycerol by a hyper producing mutant of Clostridium pasteurianum

Biotechnological products and process engineering

Abstract

Butanol, a four-carbon primary alcohol (C4H10O), is an important industrial chemical and has a good potential to be used as a superior biofuel. Bio-based production of butanol from renewable feedstock is a promising and sustainable alternative to substitute petroleum-based fuels. Here, we report the development of a process for butanol production from glycerol, which is abundantly available as a byproduct of biodiesel production. First, a hyper butanol producing strain of Clostridium pasteurianum was isolated by chemical mutagenesis. The best mutant strain, C. pasteurianum MBEL_GLY2, was able to produce 10.8 g l−1 butanol from 80 g l−1 glycerol as compared to 7.6 g l−1 butanol produced by the parent strain. Next, the process parameters were optimized to maximize butanol production from glycerol. Under the optimized batch condition, the butanol concentration, yield, and productivity of 17.8 g l−1, 0.30 g g−1, and 0.43 g l−1 h−1 could be achieved. Finally, continuous fermentation of C. pasteurianum MBEL_GLY2 with cell recycling was carried out using glycerol as a major carbon source at several different dilution rates. The continuous fermentation was run for 710 h without strain degeneration. The acetone–butanol–ethanol productivity and the butanol productivity of 8.3 and 7.8 g l−1 h−1, respectively, could be achieved at the dilution rate of 0.9 h−1. This study reports continuous production of butanol with reduced byproducts formation from glycerol using C. pasteurianum, and thus could help design a bioprocess for the improved production of butanol.

Keywords

Anaerobic fermentation Butanol Clostridium pasteurianum Glycerol Acetone–butanol–ethanol fermentation 

References

  1. Andrade JC, Vasconcelos I (2003) Continuous cultures of Clostridium acetobutylicum: culture stability and low-grade glycerol utilisation. Biotechnol Lett 25:121–125CrossRefGoogle Scholar
  2. Annous BA, Blaschek HP (1991) Isolation and characterization of Clostridium acetobutylicum mutants with enhanced amylolytic activity. Appl Environ Microbiol 57:2544–2548Google Scholar
  3. Biebl H (2001) Fermentation of glycerol by Clostridium pasteurianum: batch and continuous culture studies. J Ind Microbiol Biotechnol 27:18–26CrossRefGoogle Scholar
  4. Chang HN, Kim NJ, Kang J, Jeong CM, Choi JD, Fei Q, Kim BJ, Kwon S, Lee SY, Kim J (2010) Multi-stage high cell continuous fermentation for high productivity and titer. Bioprocess Biosyst Eng 34:419–431CrossRefGoogle Scholar
  5. Colomban A, Roger L, Boyaval P (1993) Production of propionic acid from whey permeate by sequential fermentation, ultrafiltration, and cell recycling. Biotechnol Bioeng 42:1091–1098CrossRefGoogle Scholar
  6. Connor MR, Cann AF, Liao JC (2010) 3-Methyl-1-butanol production in Escherichia coli: random mutagenesis and two-phase fermentation. Appl Microbiol Biotechnol 86:1155–1164CrossRefGoogle Scholar
  7. da Silva GP, Mack M, Contiero J (2009) Glycerol: a promising and abundant carbon source for industrial microbiology. Biotechnol Adv 27:30–39CrossRefGoogle Scholar
  8. Dabrock B, Bahl H, Gottschalk G (1992) Parameters affecting solvent production by Clostridium pasteurianum. Appl Environ Microbiol 58:1233–1239Google Scholar
  9. Durre P (2007) Biobutanol: an attractive biofuel. Biotechnol J 2:1525–1534CrossRefGoogle Scholar
  10. Ezeji TC, Qureshi N, Blaschek HP (2003) Production of acetone, butanol and ethanol by Clostridium beijerinckii BA101 and in situ recovery by gas stripping. World J Microbiol Biotechnol 19:595–603CrossRefGoogle Scholar
  11. Ezeji TC, Qureshi N, Blaschek HP (2009) Process for continuous solvent production. USA Patent, US2009/0162912 A1Google Scholar
  12. Green EM (2011) Fermentative production of butanol—the industrial perspective. Curr Opin Biotechnol 22:337–343CrossRefGoogle Scholar
  13. Jang YS, Lee J, Malaviya A, Seung DY, Cho JH, Lee SY (2011) Butanol production from renewable biomass: rediscovery of metabolic pathways and metabolic engineering. Biotechnol J. doi:10.1002/biot.201100059
  14. Johnson DT, Taconi KA (2007) The glycerin glut: options for the value-added conversion of crude glycerol resulting from biodiesel production. Environ Prog 26:338–348CrossRefGoogle Scholar
  15. Kwon S, Yoo IK, Lee WG, Chang HN, Chang YK (2001) High-rate continuous production of lactic acid by Lactobacillus rhamnosus in a two-stage membrane cell-recycle bioreactor. Biotechnol Bioeng 73:25–34CrossRefGoogle Scholar
  16. Lee J, Yun H, Feist AM, Palsson BO, Lee SY (2008a) Genome-scale reconstruction and in silico analysis of the Clostridium acetobutylicum ATCC 824 metabolic network. Appl Microbiol Biotechnol 80:849–862CrossRefGoogle Scholar
  17. Lee JY, Jang YS, Lee J, Papoutsakis ET, Lee SY (2009) Metabolic engineering of Clostridium acetobutylicum M5 for highly selective butanol production. Biotechnol J 4:1432–1440CrossRefGoogle Scholar
  18. Lee SY, Lee DY, Kim TY (2005) Systems biotechnology for strain improvement. Trends Biotechnol 23:349–358CrossRefGoogle Scholar
  19. Lee SY, Park JH, Jang SH, Nielsen LK, Kim J, Jung KS (2008b) Fermentative butanol production by Clostridia. Biotechnol Bioeng 101:209–228CrossRefGoogle Scholar
  20. Lütke-Eversloh T, Bahl H (2011) Metabolic engineering of Clostridium acetobutylicum: recent advances to improve butanol production. Curr Opin Biotechnol. doi:10.1016/j.copbio.2011.01.011
  21. Meynial-Salles I, Dorotyn S, Soucaille P (2008) A new process for the continuous production of succinic acid from glucose at high yield, titer, and productivity. Biotechnol Bioeng 99:129–135CrossRefGoogle Scholar
  22. Park JH, Lee SY, Kim TY, Kim HU (2008) Application of systems biology for bioprocess development. Trends Biotechnol 26:404–412CrossRefGoogle Scholar
  23. Qureshi N, Blaschek HP (2001) ABE production from corn: a recent economic evaluation. J Ind Microbiol Biotechnol 27:292–297CrossRefGoogle Scholar
  24. Qureshi N, Ezeji TC (2008) Butanol, ‘a superior biofuel’ production from agricultural residues (renewable biomass): recent progress in technology. Biofuels Bioprod Biorefining 2:319–330CrossRefGoogle Scholar
  25. Schlote D, Gottschalk G (1986) Effect of cell recycle on continuous butanol–acetone fermentation with Clostridium acetobutylicum under phosphate limitation. Appl Microbiol Biotechnol 24:1–5CrossRefGoogle Scholar
  26. Taconi KA, Venkataramanan KP, Johnson DT (2009) Growth and solvent production by Clostridium pasteurianum ATCC® 6013™ utilizing biodiesel-derived crude glycerol as the sole carbon source. Environ Prog Sustain Energy 28:100–110CrossRefGoogle Scholar
  27. 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 120:197–206CrossRefGoogle Scholar
  28. Vasconcelos I, Girbal L, Soucaille P (1994) Regulation of carbon electron flow in Clostridium acetobutylicum grown in chemostat culture at neutral pH on mixtures of glucose and glycerol. J Bacteriol 176:1443–1450Google Scholar
  29. 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

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  1. 1.Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 Program), BioProcess Engineering Research Center, Center for Systems and Synthetic Biotechnology, Institute for the BioCenturyKAISTDaejeonRepublic of Korea
  2. 2.Bioinformatics Research CenterKAISTDaejeonRepublic of Korea
  3. 3.BioFuelChemDaejeonRepublic of Korea

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