Three strains of Clostridium thermocellum obtained from various sources were found to have nearly identical deoxyribonucleic acid guanosine plus cytosine contents that ranged from 38.1–39.5 mole-%. All strain examined fermented only cellulose and cellulose derivatives, but not glucose, or xylose or other sugars. The principal cellulose fermentation products were ethanol, lactate, acetate, hydrogen and carbon dioxide. Growth of C. thermocellum on cellulose resulted in the production of extracellular cellulase that was non-oxygen labile, was thermally stable at 70° C for 45 min and adsorbed strongly on cellulose. Production of cellulase during fermentation correlated linearly with growth and cellulose degradation. Both the yield and specific activity of crude cellulase varied considerably with the specific growth substrates. Highest cellulase yield was obtained when grown on native cellulose, α-cellulose and low degree of polymerization cellulose but not carboxymethylcellulose or other carbohydrate sources. Cellulase activity was not detected when cells were grown on cellobiose. Crude extracellular protein preparations lacked proteolytic and cellobiase activity. The pH and temperafure optima for endoglucanase activity were 5.2 and 65° C, respectively, while that of the exoglucanase activity were 5.4 and 64° C, respectively. The specific activity at 60° c for exoglucanase and endoglucanase of crude cellulase obtained from cells grown on cellulose (MN 300) was 3.6 μmoles reducing sugar equivalents released per h (unit)/mg of protein and 1.5 μmole reducing sugar equivalent released per min (unit)/mg of protein, respectively. The yield of endoglucanase was 125 units per g of cellulose MN 300 degraded and that of exoglucanase was 300 units per g of cellulose MN 300 degraded. Glucose and cellobiose were the hydrolytic end products of crude cellulase action on cellulose, cellotraose and cellotriose in vitro.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Price excludes VAT (USA)
Tax calculation will be finalised during checkout.
Alexander, J. K.: Cellobiose phosphorylase from Clostridium thermocellum. In: Methods in enzymology, Vol. 28 (V. Ginsburg, ed.), pp. 944–948. New York: Academic Press 1972a
Alexander, J. K.: Cellodextrin phosphorylase from Clostridium thermocellum. In: Methods in enzymology, Vol. 28 (V. Ginsburg, ed.), pp. 948–953. New York: Academic Press 1972b
Bailey, M., Enari, F. M., Linkó, M.: Symposium on enzymatic hydrolysis of cellulose, Aulanko, Finland, 12–14 March 1975. The Finnish National Fund for Research and Development Helsinki, Finland (1975)
Berghem, L. E. R., Pettersson, L. G.: The mechanism of enzymatic cellulose degradation. Isolation and some properties of a β-glucosidase from Trichoderma viride. Europ. J. Biochem. 46, 295–305 (1974)
Bryant, M. P.: Commentary on the Hungate technique for culture of anaerobic bacteria. Amer. J. clin. Nutr. 25, 1324–1328 (1972)
Cooney, C. L., Ackerman, R. A.: Thermophilic anaerobic digestion of cellulosic waste. Europ. J. appl. Microbiol. 2, 65–72 (1975)
Cooney, C. L., Wise, D. L.: Thermophilic anaerobic digestion of solid waster for fuel gas production. Biotechnol. Bioeng. 17, 1119–1135 (1975)
Dawson, R. W. C., Elliott, D. C., Elliott, W. H., Jones, K. M.: Data for biochemical research, 2nd ed., p. 485. New York: Oxford Univ. Press 1972
DeLey, J.: Reexamination of the association between melting point, buoyant density and chemical base composition of deoxyribonucleic acid. J. Bact. 101, 738–754 (1970)
Enebo, L., Lundin, H.: On three bacteria connected with thermophilic cellulose fermentation. Physiol. Plant. 4, 652–660 (1951)
Feather, M. S., Harris, J. F.: Chromatographic isolation of some cellulose oligosaccharides. U.S. Forest Service Res. Note. FPL-0161. 8. p (1967)
Hungate, R. E.: A roll tube method for cultivation of strict anaerobes. In: Methods in microbiology, Vol. 3B (J. R. Norris, D. W. Ribbons, eds.), p. 117. New York-London: Academic Press 1969.
Kaustinen, H. M., Kaustinen, O. A., Swenson, H. A.: Selective acetolysis of cellulose to low degree of polymerization. Carbohyd. Res. 11, 267–268 (1969)
Lee, B. H., Blackburn, T. H.: Cellulase production by a thermophilic Clostridium species. Appl. Microbiol. 30, 346–353 (1975)
Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. I.: Protein measurement with the Folin Phenol Reagent. J. biol. Chem. 193, 265–275 (1951)
Marmur, J.: A procedure for isolation of deoxyribonucleic acid from microorganisms. J. molec. Biol. 3, 208–218 (1961)
McBee, R. H.: The culture and physiology of a thermophilic cellulose-fermenting bacterium. J. Bact. 56, 653–663 (1948)
McBee, R. H.: The anaerobic thermophilic cellulolytic bacteria. Bact. Rev. 14, 51–63 (1950)
Miller G. L., Blum, R., Glennon, W. E., Burton, A. L.: Measurement of carboxymethylcellulase activity. Analyt. Biochem. 1, 127–132 (1960).
Millett, M. A., Moore, W. E., Saeman, J. F.: Techniques for quantiative thin layer chromatography. Analyt. Chem. 36, 491–494 (1964)
Moore, S., Stein, W. H.: A modified minhydrin reagent for the photometric determination of amino acids and related compounds. J. biol. Chem. 211, 907–913 (1954)
Nelson, N.: A photometric adaptation of the Somogyi method for the determination of glucose. J. biol. Chem. 153, 375–380 (1944)
Norkrans, B.: Studies of β-glucoside and cellulose splitting enzymes from Polyporus amnosus Fr. Physiol. Plant. 10, 198–214 (1957)
Patni, N. J., Alexander, J. K.: Catabolism of fructose and mannitol in Clostridium thermocellum: Presence of phosphoenol-pyruvate: fructose phosphotransferase, fructose 1-phosphate kinase, phosphoenolpyruvate: mannitol phosphotransferase, and mannitol 1-phosphate dehydrogenase in cell extracts. J. Bact. 105, 226–231 (1971a)
Patni, N. J., Alexander, J. K.: Utilization of glucose by Clostridium thermocellum: Presence of glucosinase and other glycolytic enzymes in cell extracts. J. Bact. 105, 220–225 (1971b)
Pfeffer, J. T.: Temperature effects on anaerobic fermentation of domestic wastes. Biotechnol. Bioeng. 16, 771–787 (1974)
Reese, E. T.: A microbiological progress report. Enzymatic hydrolysis of cellulose. Appl. Microbiol. 4, 39–45 (1956)
Romanelli, R. A., Houston, C. W., Barnett, S. M.: Studies on thermophilic cellulolytic fungi. Appl. Microbiol. 30, 276–281 (1975)
Somogyi, M.: Notes on sugar determinations. J. biol. Chem. 195, 19–23 (1952)
Stutzenberger, F. J., Kaufman, A. J., Lossin, R. D.: Cellulolytic activity in municipal waste composting. Canad. J. Microbiol. 16, 553–560 (1970)
Viljoen, J. A., Fred, E. B., Peterson, W. H.: The fermentation of cellulose by thermophilic bacteria. J. Agric. Sci. 16, 1–17 (1926)
Weimer, P. J., Zeikus, J. G.: Fermentation of cellulose and cellobiose by C. thermocellum in the absence and presence of Methanobacterium thermoautotrophicum. Appl. Environ. Microbiol. 33, 289–297 (1977)
Wilke, C. R.: Cellulose as a chemical and energy resource. Biotech. & Bioeng. Symposium No. 5. New York: John Wiley & Sons 1975
About this article
Cite this article
Ng, T.K., Weimer, P.J. & Zeikus, J.G. Cellulolytic and physiological properties of Clostridium thermocellum . Arch. Microbiol. 114, 1–7 (1977). https://doi.org/10.1007/BF00429622