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. 1980 Oct 1;191(1):83–94. doi: 10.1042/bj1910083

Purification and properties of the cellulases from the thermophilic fungus Thermoascus aurantiacus.

C C Tong, A L Cole, M G Shepherd
PMCID: PMC1162184  PMID: 6781492

Abstract

Three cellulases and a beta-glucosidase were purified from the culture filtrate of the thermophilic fungus Thermoascus aurantiacus. The isolated enzymes were all homogeneous on polyacrylamide-disc-gel electrophoresis. Data from chromatography on Bio-Gel P-60 and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis indicated mol.wts. of 87000 (beta-glucosidase), 78000 (cellulase I), 49000 (cellulase II) and 34000 (cellulase III); the carbohydrate contents of the enzymes were 33.0, 5.5, 2.6 and 1.8% (w/w) respectively. Although the three purified cellulases were active towards filter paper, only cellulases I and III were active towards CM(carboxymethyl)-cellulose. Cellulase I was also active towards yeast glucan. The Km and catalytic-centre-activity values for the enzymes were as follows; 0.52 mumol/ml and 6.5 X 10(4) for beta-glucosidase on p-nitrophenyl beta-D-glucoside, 3.9 mg/ml and 6.3 for cellulase I on CM-cellulose, 1.2 mg/ml and 1.1 for cellulase I on yeast glucan, 35.5 mg/ml and 0.34 for cellulase II on filter paper, and 1.9 mg/ml and 33 for cellulase III on CM-cellulose.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andrews P. Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochem J. 1964 May;91(2):222–233. doi: 10.1042/bj0910222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berghem L. E., Pettersson L. G. The mechanism of enzymatic cellulose degradation. Isolation and some properties of a beta-glucosidase from Trichoderma viride. Eur J Biochem. 1974 Jul 15;46(2):295–305. doi: 10.1111/j.1432-1033.1974.tb03621.x. [DOI] [PubMed] [Google Scholar]
  3. Berghem L. E., Pettersson L. G. The mechanism of enzymatic cellulose degradation. Purification of a cellulolytic enzyme from Trichoderma viride active on highly ordered cellulose. Eur J Biochem. 1973 Aug 1;37(1):21–30. doi: 10.1111/j.1432-1033.1973.tb02952.x. [DOI] [PubMed] [Google Scholar]
  4. Eisenthal R., Cornish-Bowden A. The direct linear plot. A new graphical procedure for estimating enzyme kinetic parameters. Biochem J. 1974 Jun;139(3):715–720. doi: 10.1042/bj1390715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Eriksson K. E., Hollmark B. H. Kinetic studies of the action of cellulase upon sodium carboxymethyl cellulose. Arch Biochem Biophys. 1969 Sep;133(2):233–237. doi: 10.1016/0003-9861(69)90450-0. [DOI] [PubMed] [Google Scholar]
  6. Eriksson K. E., Pettersson G. Studies on cellulolytic enzymes. V. Some structural properties of the cellulase from Penicillium notatum. Arch Biochem Biophys. 1968 Mar 20;124(1):160–166. doi: 10.1016/0003-9861(68)90316-0. [DOI] [PubMed] [Google Scholar]
  7. Fergus C. L. The cellulolytic activity of thermophilic fungi and Actinomycetes. Mycologia. 1969 Jan-Feb;60(1):120–129. [PubMed] [Google Scholar]
  8. HALLIWELL G. HYDROLYSIS OF FIBROUS COTTON AND REPRECIPITATED CELLULOSE BY CELLULOLYTIC ENZYMES FROM SOIL MICRO-ORGANISMS. Biochem J. 1965 Apr;95:270–281. doi: 10.1042/bj0950270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. HASH J. H., KING K. W. On the nature of the beta-glucosidases of Myrothecium verrucaria. J Biol Chem. 1958 May;232(1):381–393. [PubMed] [Google Scholar]
  10. Hurst P. L., Nielsen J., Sullivan P. A., Shepherd M. G. Purification and properties of a cellulase from Aspergillus niger. Biochem J. 1977 Jul 1;165(1):33–41. doi: 10.1042/bj1650033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Li L. H., Flora R. M., King K. W. Individual roles of cellulase components derived from Trichoderma viride. Arch Biochem Biophys. 1965 Aug;111(2):439–447. doi: 10.1016/0003-9861(65)90207-9. [DOI] [PubMed] [Google Scholar]
  12. ORNSTEIN L. DISC ELECTROPHORESIS. I. BACKGROUND AND THEORY. Ann N Y Acad Sci. 1964 Dec 28;121:321–349. doi: 10.1111/j.1749-6632.1964.tb14207.x. [DOI] [PubMed] [Google Scholar]
  13. PETTERSSON G., COWLING E. B., PORATH J. Studies on celluloytic enzymes. I. Isolation of a low-molecular-weight cellulase from Polyporus versicolor. Biochim Biophys Acta. 1963 Jan 8;67:1–8. doi: 10.1016/0006-3002(63)91791-8. [DOI] [PubMed] [Google Scholar]
  14. Reisner A. H., Nemes P., Bucholtz C. The use of Coomassie Brilliant Blue G250 perchloric acid solution for staining in electrophoresis and isoelectric focusing on polyacrylamide gels. Anal Biochem. 1975 Apr;64(2):509–516. doi: 10.1016/0003-2697(75)90461-3. [DOI] [PubMed] [Google Scholar]
  15. Romanelli R. A., Houston C. W., Barnett S. M. Studies on thermophilic cellulolytic fungi. Appl Microbiol. 1975 Aug;30(2):276–281. doi: 10.1128/am.30.2.276-281.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rosenberg S. L. Temperature and pH optima for 21 species of thermophilic and thermotolerant fungi. Can J Microbiol. 1975 Oct;21(10):1535–1540. doi: 10.1139/m75-225. [DOI] [PubMed] [Google Scholar]
  17. SELBY K., MAITLAND C. C. THE FRACTIONATION OF MYROTHECIUM VERRUCARIA CELLULASE BY GEL FILTRATION. Biochem J. 1965 Mar;94:578–583. doi: 10.1042/bj0940578. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. SMOGYI M. Notes on sugar determination. J Biol Chem. 1952 Mar;195(1):19–23. [PubMed] [Google Scholar]
  19. Selby K., Maitland C. C. The cellulase of Trichoderma viride. Separation of the components involved in the solubilization of cotton. Biochem J. 1967 Sep;104(3):716–724. doi: 10.1042/bj1040716. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Stutzenberger F. J. Cellulase production by Thermomonospora curvata isolated from municipal solid waste compost. Appl Microbiol. 1971 Aug;22(2):147–152. doi: 10.1128/am.22.2.147-152.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Weber K., Pringle J. R., Osborn M. Measurement of molecular weights by electrophoresis on SDS-acrylamide gel. Methods Enzymol. 1972;26:3–27. doi: 10.1016/s0076-6879(72)26003-7. [DOI] [PubMed] [Google Scholar]
  22. Wood T. M., McCrae S. I. The purification and properties of the C 1 component of Trichoderma koningii cellulase. Biochem J. 1972 Aug;128(5):1183–1192. doi: 10.1042/bj1281183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wood T. M., Phillips D. R. Another source of cellulase. Nature. 1969 Jun 7;222(5197):986–987. doi: 10.1038/222986b0. [DOI] [PubMed] [Google Scholar]
  24. Wood T. M. The cellulase of Fusarium solani. Resolution of the enzyme complex. Biochem J. 1969 Nov;115(3):457–464. doi: 10.1042/bj1150457. [DOI] [PMC free article] [PubMed] [Google Scholar]

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