Applied Microbiology and Biotechnology

, Volume 29, Issue 6, pp 528–535

Ethanol production by Clostridium thermocellum grown on hydrothermally and organosolv-pretreated lignocellulosic materials

  • H. F. Hörmeyer
  • P. Tailliez
  • J. Millet
  • H. Girard
  • G. Bonn
  • O. Bobleter
  • J. -P. Aubert
Biotechnology

Summary

Two strains of the thermophilic anaerobe Clostridium thermocellum, the wild type NCIB 10682 and its ethanol-hyperproductive mutant 647, were tested for their ability to grow on natural lignocellulosic materials (poplar wood, wheat straw) which had been pretreated by either hydrothermolysis or an organosolv process. For both materials and both strains, the dependencies of substrate accessibility on the pretreatment temperature were established in terms of cellulose hydrolysis and of product formation.

In addition to the non-pH-controlled shake flask assays, in vitro experiments with cell-free culture supernatant and in vivo cellulolyses under pH regulation in a laboratory fermenter indicated that lignocellulosics pretreated at approx. 230°C were degraded efficiently by the Clostridium strains investigated.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Bayer EA, Setter E, Lamed R (1985) Organization and distribution of the cellulosome in Clostridium thermocellum. J Bacteriol 163: 552–559Google Scholar
  2. Bobleter O, Bonn G, Prutsch W (1987) Production of hemicellulose and pulp by hydrothermal processes (Presented at the Conference on Non-Conventional Pulping and Bleaching, May 4–5, 1987, Hamburg/FRG)Google Scholar
  3. Bobleter O, Pape G (1968) Verfahren zum Abbau von Holz, Rinde oder anderen Pflanzenmaterialien. Aust Pat no. 263.661Google Scholar
  4. Bonn G, Concin R, Bobleter O (1983) Hydrothermolysis — a new process for the utilization of biomass. Wood Sci Technol 17: 195–202Google Scholar
  5. Bonn G, Hörmeyer HF, Bobleter O (1987) Hydrothermal and organosolv pretreatments of poplar wood and wheat straw for saccharification by a Trichoderma viride cellulase. Wood Sci Technol 21: 179–185Google Scholar
  6. Cowling EB, Kirk TK (1976) Properties of cellulose and lignocellulosic materials as substrates for enzymic conversion processes. Biotechnol Bioeng Symp 6: 95–123Google Scholar
  7. Edel E, Feckl J (1985) Studies of soluble by-products resulting from organosolv pulping. Report on contract no. BOS-008-D, presented at CEC Contractors' Meeting (Friedrichshafen/FRG)Google Scholar
  8. Fan LT, Lee Y-H, Beardmore DH (1980) Mechanism of the enzymic hydrolysis of cellulose: effects of the major structural features of cellulose on enzymic hydrolysis. Biotechnol Bioeng 22: 177–199Google Scholar
  9. Hörmeyer HF, Bonn G, Kim DW, Bobleter O (1987) Enzymatic saccharification of cellulosic materials after hydrothermolysis and organosolv pretreatments. J Wood Chem Technol 7: 269–283Google Scholar
  10. Johnson EA, Demain AL (1984) Probable involvement of sulfhydryl groups and a metal as essential components of the cellulase of Clostridium thermocellum. Arch Microbiol 137: 135–138Google Scholar
  11. Johnson EA, Sakajoh M, Halliwell G, Madia A, Demain AL (1982) Saccharification of complex cellulosic substrates by the cellulase system from Clostridium thermocellum. Appl Environ Microbiol 43: 1125–1132Google Scholar
  12. Joliff G, Béguin P, Juy M, Millet J, Ryter A, Poljak R, Aubert J-P (1986) Isolation, crystallization and properties of a new cellulase of Clostridium thermocellum overproduced in Escherichia coli. Bio/Technology 4: 896–900Google Scholar
  13. Kleinert TN (1974) Organosolv pulping with aqueous alcohol. Tappi 57: 99–102Google Scholar
  14. Lynd LR, Grethlein HE (1987) Hydrolysis of dilute acid pretreated mixed hardwood and purified microcrystalline cellulose by cell-free broth from Clostridium thermocellum. Biotechnol Bioeng 29: 92–100Google Scholar
  15. Millet J, Pétré D, Béguin P, Raynaud O, Aubert J-P (1985) Cloning of ten distinct DNA fragments of Clostridium thermocellum coding for cellulases. FEMS Microbiol Lett 29: 145–149Google Scholar
  16. Ng TK, Weimer PJ, Zeikus JG (1977) Cellulolytic and physiological properties of Clostridium thermocellum. Arch Microbiol 114: 1–7Google Scholar
  17. Pecina R, Bonn G, Burtscher E, Bobleter O (1984) High-performance liquid chromatographic elution behaviour of alcohols, aldehydes, ketones, organic acids and carbohydrates on a strong cation-exchange stationary phase. J Chromatogr 284: 245–258Google Scholar
  18. Rapp P, Grote E, Wagner F (1981) Formation and location of 1,4-β-glucanases and 1,4-β-glucosidases from Penicillium janthinellum. Appl Environ Microbiol 41: 857–866Google Scholar
  19. Reese ET, Mandels M (1984) Rolling with the times: production and applications of Trichoderma reesei cellulase. Annu Rep Ferment Processes 7: 1–20Google Scholar
  20. Romaniec MPM, Clarke NG, Hazelwood GP (1987) Molecular cloning of Clostridium thermocellum DNA and the expression of further novel endo-β-1,4-glucanase genes in Escherichia coli. J Gen Microbiol 133: 1297–1307Google Scholar
  21. Schwarz W, Bronnenmeier K, Staudenbauer WL (1985) Molecular cloning of Clostridium thermocellum genes involved in β-glucan degradation in bacteriophage lambda. Biotechnol Lett 7: 859–864Google Scholar
  22. Schwarz WH, Grabnitz G, Staudenbauer WL (1986) Properties of a Clostridium thermocellum endoglucanase produced in Escherichia coli. Appl Environ Microbiol 51: 1293–1299Google Scholar
  23. Sedmak JJ, Grossberg SE (1977) A rapid, sensitive and versatile assay for protein using Coomassie brilliant blue G250. Anal Biochem 79: 544–552Google Scholar
  24. Wang DIC, Avgerinos GC, Biocic I, Wang SD, Fang HY (1983) Ethanol from cellulosic biomass. Phil Trans R Soc London, B 300: 323–333Google Scholar
  25. Weimer PJ, Weston WM (1985) Relationship between the fine structure of native cellulose and cellulose degradability by the cellulase complexes of Trichoderma reesei and Clostridium thermocellum. Biotechnol Bioeng 27: 1540–1547Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • H. F. Hörmeyer
    • 1
  • P. Tailliez
    • 2
  • J. Millet
    • 2
  • H. Girard
    • 2
  • G. Bonn
    • 1
  • O. Bobleter
    • 1
  • J. -P. Aubert
    • 2
  1. 1.Institut für Radiochemie der Universität InnsbruckInnsbruckAustria
  2. 2.Departement des Biotechnologies, Unité de Physiologie Cellulaire, 28Institut PasteurParis Cédex 15France

Personalised recommendations