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. 1981 Apr;41(4):924–931. doi: 10.1128/aem.41.4.924-931.1981

Separation and Some Properties of Two Intracellular β-Glucosidases of Sporotrichum (Chrysosporium) thermophile

Hans-Peter Meyer 1,†, Giorgio Canevascini 1
PMCID: PMC243835  PMID: 16345755

Abstract

Intracellular, inducible β-glucosidase from the cellulolytic fungus Sporotrichum (Chrysosporium) thermophile (ATCC 42464) was fractionated by gel chromatography or isoelectric focusing into components A and B. Enzyme A (molecular weight 440,000) had only aryl-β-glucosidase activity, whereas enzyme B (molecular weight 40,000) hydrolyzed several β-glucosides but had only low activity against o-nitrophenyl-β-d-glucopyranoside (ONPG). Both enzymes had temperature optima of about 50°C. The pH optimum was 5.6 for enzyme A and 6.3 for enzyme B, respectively. The Km (ONPG) value for enzyme A was 0.5 mM, and the corresponding values for enzyme B were 0.18 mM (ONPG) and 0.28 mM (cellobiose). Enzyme B, when tested with ONPG, showed substrate inhibition at a substrate concentration above 0.4 mM which could be released by cellobiitol and other alditols. Enzyme A was isoelectric at pH 4.48, and enzyme B was isoelectric at pH 4.64. Several inhibitors were tested for their action on the activity of enzymes A and B. Both enzymes were found to be concomitantly induced in cultures with either cellobiose or cellulose as carbon source.

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Selected References

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  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. Aït N., Creuzet N., Cattanéo J. Characterization and purification of thermostable beta-glucosidase from Clostridium thermocellum. Biochem Biophys Res Commun. 1979 Sep 27;90(2):537–546. doi: 10.1016/0006-291x(79)91269-5. [DOI] [PubMed] [Google Scholar]
  3. CROOK E. M., STONE B. A. Formation of oligosaccharides during the enzymic hydrolysis of beta-glucosides. Biochem J. 1953 Jul 17;55(320TH):xxv–xxv. [PubMed] [Google Scholar]
  4. Cortat M., Matile P., Wiemken A. Isolation of glucanase-containing vesicles from budding yeast. Arch Mikrobiol. 1972;82(3):189–205. doi: 10.1007/BF00412191. [DOI] [PubMed] [Google Scholar]
  5. DUERKSEN J. D., HALVORSON H. Purification and properties of an inducible beta-glucosidase of yeast. J Biol Chem. 1958 Nov;233(5):1113–1120. [PubMed] [Google Scholar]
  6. Deshpande V., Eriksson K. E., Pettersson B. Production , purification and partial characterization of 1,4-beta-glucosidase enzymes from Sporotrichum pulverulentum. Eur J Biochem. 1978 Sep 15;90(1):191–198. doi: 10.1111/j.1432-1033.1978.tb12590.x. [DOI] [PubMed] [Google Scholar]
  7. Eberhart B. M., Beck R. S. Localization of the beta-glucosidases in Neurospora crassa. J Bacteriol. 1970 Feb;101(2):408–417. doi: 10.1128/jb.101.2.408-417.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. HU A. S., EPSTEIN R., HALVORSON H. O., BOCK R. M. Yeast beta-glucosidase: comparison of the physical-chemical properties of purified constitutive and inducible enzyme. Arch Biochem Biophys. 1960 Dec;91:210–218. doi: 10.1016/0003-9861(60)90492-6. [DOI] [PubMed] [Google Scholar]
  9. Han Y. W., Srinivasan V. R. Purification and characterization of beta-glucosidase of Alcaligenes faecalis. J Bacteriol. 1969 Dec;100(3):1355–1363. doi: 10.1128/jb.100.3.1355-1363.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hägerdal B., Harris H., Pye E. K. Association of beta-glucosidase with intact cells of Thermoactinomyces. Biotechnol Bioeng. 1979 Mar;21(3):345–355. doi: 10.1002/bit.260210302. [DOI] [PubMed] [Google Scholar]
  11. Inamdar A. N., Kaplan J. G. Purification and properties of an inducible beta-glucosidase of Bakers' yeast. Can J Biochem. 1966 Aug;44(8):1099–1108. doi: 10.1139/o66-127. [DOI] [PubMed] [Google Scholar]
  12. Inglin M., Feinberg B. A., Loewenberg J. R. Partial purification and characterization of a new intracellular beta-glucosidase of Trichoderma reesei. Biochem J. 1980 Feb 1;185(2):515–519. doi: 10.1042/bj1850515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Legler G. Untersuchungen zum Wirkungsmechanismus glykosidspaltender Enzyme, II. Isolierung und enzymatische Eigenschaften von zwei beta-Glucosidasen aus Aspergillus wentii. Hoppe Seylers Z Physiol Chem. 1967 Nov;348(11):1359–1366. [PubMed] [Google Scholar]
  15. 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]
  16. Lusis A. J., Becker R. R. The beta-glucosidase system of the thermophilic fungus Chaetomium thermophile var. coprophile n. var. Biochim Biophys Acta. 1973 Nov 2;329(1):5–16. doi: 10.1016/0304-4165(73)90003-2. [DOI] [PubMed] [Google Scholar]
  17. Roth W. W., Srinivasan V. R. Affinity chromatographic purification of beta-glucosidase of Candida gulliermondii. Prep Biochem. 1978;8(1):57–71. doi: 10.1080/00327487808068218. [DOI] [PubMed] [Google Scholar]
  18. Rudick M. J., Elbein A. D. Glycoprotein enzymes secreted by Aspergillus fumigatus: purification and properties of a second beta-glucosidase. J Bacteriol. 1975 Oct;124(1):534–541. doi: 10.1128/jb.124.1.534-541.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sternberg D., Vijayakumar P., Reese E. T. beta-Glucosidase: microbial production and effect on enzymatic hydrolysis of cellulose. Can J Microbiol. 1977 Feb;23(2):139–147. doi: 10.1139/m77-020. [DOI] [PubMed] [Google Scholar]
  20. Umezurike G. M. Kinetic analysis of the mechanism of action of beta-glucosidase from Botryodiplodia theobromae Pat. Biochim Biophys Acta. 1975 Jul 27;397(1):164–178. doi: 10.1016/0005-2744(75)90190-4. [DOI] [PubMed] [Google Scholar]
  21. Umezurike G. M. The cellulolytic enzymes of Botryodiplodia theobromae Pat. Separation and characterization of cellulases and beta-glucosidases. Biochem J. 1979 Jan 1;177(1):9–19. doi: 10.1042/bj1770009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Umezurike G. M. The purification and properties of extracellular beta-glucosidase from Botryodiplodia theobromae Pat. Biochim Biophys Acta. 1971 Feb 10;227(2):419–428. doi: 10.1016/0005-2744(71)90073-8. [DOI] [PubMed] [Google Scholar]
  23. Umezurike G. M. The subunit structure of beta-glucosidase from Botryodiplodia theobromae Pat. Biochem J. 1975 Feb;145(2):361–368. doi: 10.1042/bj1450361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. de Gussem R. L., Aerts G. M., Claeyssens M., de Bruyne C. K. Purification and properties of an induced beta-D-glucosidase from stachybotrys atra. Biochim Biophys Acta. 1978 Jul 7;525(1):142–153. doi: 10.1016/0005-2744(78)90208-5. [DOI] [PubMed] [Google Scholar]

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