Export file:


  • RIS(for EndNote,Reference Manager,ProCite)
  • BibTex
  • Text


  • Citation Only
  • Citation and Abstract

Switchgrass (Panicum virgatum) fermentation by Clostridium thermocellum and Clostridium saccharoperbutylacetonicum sequential culture in a continuous flow reactor

1 University of Kentucky, Department of Biosystems and Agricultural Engineering, Lexington, Kentucky, USA
2 USDA, Agricultural Research Service, Forage-Animal Production Research Unit, Lexington, Kentucky, USA
3 University of Kentucky, Department of Animal and Food Sciences, Lexington, Kentucky, USA

Special Issues: Advances in Production of Biofuels

The study was conducted to evaluate fermentation by Clostridium thermocellum and C. saccharoperbutylacetonicum in a continuous-flow, high-solids reactor. Liquid medium was continuously flowed through switchgrass (2 mm particle size) at one of three flow rates: 83.33 mL h1 (2 L d−1), 41.66 mL h−1 (1 L d−1), and 20.833 mL h−1 (0.5 L d−1). The cellulolytic phase was initiated by culturing C. thermocellum (63 °C, 24 h). The temperature was decreased (35) and C. saccharoperbutylacetonicum was inoculated. When metabolism decreased (96 h), the temperature was increased (63 °C; 24 h) to permit cellulosome production by C. thermocellum. The C. saccharoperbutylacetonicum was re-inoculated and the temperature returned to 35°C. The average gross production over 9 d was 1480 mg total acids (formic, acetic lactic butyric), 207 mg total solvents (acetone, butanol, ethanol), and average dry matter disappearance was 2.8 g from 25 g non-pretreated switchgrass. There was no effect of flow rate on the product formation. These results indicate that C. thermocellum can survive and produce cellulases with C. saccharoperbutylacetonicumin a continuous-flow, high-solids reactor temperature with temperature cycling.
  Article Metrics


1. Perlack R, Lynn L, Wright A, et al. (2005) Biomass as feedstock for a bioenergy and bioproducts industry: the technical feasibility of a billion ton annual supply. Available from: http://www.osti.gov/brigde.

2. Parrish D, Fike J (2005). The biology and agronomy of switchgrass for biofuels. Crit rev plant sci 24: 423-459.    

3. Demain A, Newcomb M, Wu J (2005) Cellulase, clostridia, and ethanol. Microbiolmolbiol rev 69: 124-54.

4. Qureshi N, Ezeji T (2008) Butanol, ‘a superior biofuel’ production from agricultural residues (renewable biomass): recent progress in technology. Biofuel bioprodbiores2: 319-330.

5. Hongo M (1960) Process for producing butanol by fermentation.US Patent 2945786.

6. Jones D, Keis S (1995) Origins and relationships of industrial solvent-producing clostridial strains. FEMS microbiol rev 17: 223-232.    

7. Thang V, Kanda K, KobayanshI G (2010) Production of Acetone-Butanol-Ethanol (ABE) in direct fermentation of cassava by Clostridium saccharoperbutylacetonicum N1-4. Applbiochembiotechnol 161: 157-170.

8. Al-Shorgani NK, Kalil MS, Yusoff W (2011) The effect of different carbon sources on butanol production using Clostridium saccharoperbutylacetonicum N1-4. Biotechnol 10: 280-285.    

9. Al-Shorgani N, Kalil M, Yusoff W (2012) Biobutanol production from rice bran and de-oiled rice bran by Clostridium saccharoperbutylacetonicum N1-4. Bioprocess biosysteng35: 817-826.

10. McBee R (1954) The characteristics of Clostridium thermocellum. J Bacteriol 67: 505-506.

11. Lynd L, Weimer P, van Zyl W, et al. (2002) Microbial cellulose utilization fundamentals and biotechnology. Microbiolmolbiol rev 66: 506-577.

12. Lynd L, Van Zyl W, McBride J, et al. (2005) Consolidated bioprocessing of cellulosic biomass: An update. Curr opin biotechnol 16: 577-583.    

13. Flythe M, Elía N, Schmal M, et al. (2015) Switchgrass (Panicumvirgatum) fermentation by Clostridium thermocellum and Clostridium beijerinckii sequential culture: effect of feedstock particle size on gas production. Advmicrob 5: 311-316.

14. Chinn M, Nokes S, Strobel H (2006) Screening of thermophilic anaerobic bacteria for solid substrate cultivation on lignocellulosic substrates. Biotechnolprog 22: 53-59.

15. Dharmagadda V, Nokes S, Strobel H, et al. (2010) Investigation of the metabolic inhibition observed in solid substrate cultivation of Clostridium thermocellum on cellulose. Bioresourcetechnol101: 6039‐6044.

16. Chinn M, Nokes S, Strobel H (2007) Influence of process conditions on end product formation from Clostridium thermocellum 27405 in solid substrate cultivation on paper pulp sludge. Bioresourcetechnol98: 2184‐2193.

17. Selig M, Hsieh C, Thygesen I, et al. (2012) Considering water availability and the effect of solute concentration on high solids saccharification of lignocellulosic biomass. Biotechnolprog 28: 1478-1490.

18. Yao W, Nokes S (2014) First proof of concept of sustainable metabolite production from high solids fermentation of lignocellulosic biomass using a bacterial co-culture and cycling flush system. Bioresourcetechnol 173: 216-223.    

19. Cotta M, Russell J (1982) Effects of peptides and amino acids on efficiency of rumen bacterial protein synthesis in continuous culture. J dairy sci65: 226-234.

20. Sun Y, Cheng J (2002) Hydrolysis of lignocellulosic materials for ethanol production: A review. Bioresourcetechnol 83: 1-11    

21. Ng T, Weimer P, Zeikus J (1977) Cellulolytic and physiological properties of Clostridium thermocellum. Arch microbiol 114:1-7.    

22. Bayer E, Belaich J, Shoham Y, et al. (2004) The cellulosomes: multienzymatic machines for degradation of plant cell wall polysaccharides. Annu rev microbiol 58: 521-554.    

23. Yu E, Chan M, Saddler J (1985) Butanol production from cellulosic substrates by sequential co-culture of Clostridium thermocellum and C. acetobutylicum. Biotech letters 7: 509-514.    

24. Ni Y, Sun ZH (2009) Recent progress on industrial fermentative production of acetone-butanol-ethanol by Clostridium acetobutylicum in China. Appl microbiol biotechnol 83: 415-423.    

25. Ezeji T, Qureschi N, Blaschek H (2004) Butanol fermentation research: upstream and downstream manipulations. Chemrec 4: 305-314.

26. Kosaka T, Nakayama S, Nakayama K, et al. (2007) Characterization of the sol operon in butanol-hyperproducingClostridium saccharoperbutylacetonicum strain N1-4 and its degeneration mechanism. Biosciniotechnolbiochem 71: 58-68.

27. Yao W, Nokes S (2014) Phanerochaetechrysosporium pretreatment of biomass to enhance solvent production in subsequent bacterial solid-substrate cultivation. Biomass bioenergy 62: 100-107.

28. Kristensen J, Felby C, Jorgensen H (2009) Yield-determining factors in high solids enzymatic hydrolysis of lignocelluloses. Biotechnol biofuels 2: 11.    

29. Li H, Knutson B, Nokes S, et al. (2012) Metabolic control of Clostridium thermocellum via inhibition of hydrogenase activity and the glucose transport rate. Appl microbiol biotechnol 93: 1777-1784.    

Copyright Info: © 2016, Michael D. Flythe, et al., licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution Licese (http://creativecommons.org/licenses/by/4.0)

Download full text in PDF

Export Citation

Article outline

Show full outline
Copyright © AIMS Press All Rights Reserved