Skip to content
1887

Microbiology Society logo Microbiology

Volume 128, Issue 3

Properties of -Glucosidase Purified from Free

Abstract

A cell-bound -glucosidase (--glucoside glucohydrolase; EC 3.2.1.21) from thermocellum was purified to apparent homogeneity. A molecular weight of about 43000 gel fluration of the native enzyme on Ultrogel AcA34. A constant ratio of aryl--glucosidase and cellcouse throughout purification, similar heat stabilities, pH profiles and sensitivity to different hiibitor and competitive inhibition of the aryl--gluosidase and the suggest that the same enzyme accounts for the aryl--glucosidase and the cellobiase activities. However, the affinity for cellobiose was very much lower than for ---glucoside. The -glucosidase had maximum rates at pH 6·0 to 6·5 for both activities. The enzyme was specific for substrates with the both activities. -configuration. particulary and -1,3 and -1,2 linkages. The enzyme did not hydrolyse carboxymethylcellulose or cellulose, but hydrolysed cello-oligosaccharides. It was strongly inhibited by -glucono--lactone was sensitive to thiol reagents. When preparations of cellulase were supplemented with purified -glucosidase, glucose was the predominant product of cellulose and the rate of saccharification was increased.

  • Received:
  • Revised:
  • Published Online:
© Society for General Microbiology 1982
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-128-3-569
1982-03-01
2025-04-21
Loading full text...

Full text loading...

/deliver/fulltext/micro/128/3/mic-128-3-569.html?itemId=/content/journal/micro/10.1099/00221287-128-3-569&mimeType=html&fmt=ahah

References

  1. Aït N. 1980 Dégradation de la cellulose par Clostridium thermocellum. Caractérisation et purification d’une β-glucosidase. Ph.D. thesis Aix-Marseille, France:
     [Google Scholar]
  2. Aït N., Creuzet N., Forget P. 1979a; Partial purification of cellulase from Clostridium thermocellum. Journal of General Microbiology 113:399–402
     [Google Scholar]
  3. Aït N., Creuzet N., Cattaneo J. 1979b; Characterisation and purification of thermostable β-glucosidase from Clostridium thermocellum. Biochemical and Biophysical Research Communications 90:537–546
     [Google Scholar]
  4. Alexander J. K. 1967; Cellodextrin phosphorylase from Clostridium thermocellum. Biochimica et biophysica acta 148:808–810
     [Google Scholar]
  5. Alexander J. K. 1968; Purification and specificity of cellobiose phosphorylase from Clostridium thermocellum. Journal of Biological Chemistry 243:2899–2904
     [Google Scholar]
  6. Andrews P. 1964; Estimation of the molecular weight of proteins by Sephadex gel filtration. Biochemical Journal 91:222–233
     [Google Scholar]
  7. Berghem L. E. R., Petterson L. G. 1974; The mechanisms of enzymatic cellulose degradation. Isolation and some properties of a β-glucosidase from: Trichoderma viride. European Journal of Biochemistry 46:295–305
     [Google Scholar]
  8. Davis B. J. 1964; Disc electrophoresis. II. Method and application to human serum proteins. Annals of the New York Academy of Sciences 121:404–427
     [Google Scholar]
  9. De Gussem R. L., Aerts G. M., Claeyssens M., De Bruyne C. K. 1978; Purification and properties of an induced β-d-glucosidase from Stachybotrys atra. Biochimica et biophysica acta 525:142–153
     [Google Scholar]
  10. Dubois M., Gilles K., Hamilton J. K., Rebers P. A., Smith F. 1956; Colorimetric method for determination of sugars and related substances. Analytical Biochemistry 28:350–356
     [Google Scholar]
  11. Gordon J., Cooney C. L. 1980; Fermentation production of sugars from cellulose by Clostridium thermocellum. Abstracts of the VIth International Fermentation Symposium and VIIth International Symposium on Yeasts London, Ontario, Canada: p. 87:
     [Google Scholar]
  12. Han Y. W., Srinivasan V. R. 1969; Purification and characterization of β-glucosidase of Alcaligenes faecalis. Journal of Bacteriology 100:1353–1363
     [Google Scholar]
  13. Herr D., Baumer F., Dellweg H. 1978; Purification and properties of an extracellular β-glucosidase from Lenzites trabea. European Journal of Applied Microbiology 5:29–36
     [Google Scholar]
  14. Hirayama T., Horie S., Nagayama H., Matsuda K. 1978; Studies on cellulases of a phytopathogenic fungus, Pyricularia oryzae Cavara. II. Purification and properties of a β-glucosidase. Journal of Biochemistry 84:27–37
     [Google Scholar]
  15. Jørgensen B. B., Jørgensen O. B. 1967; Inhibition of barley malt α-glucosidase by tris(hydroxymethyl)aminomethane and erythritol. Biochimica et biophysica acta 146:167–172
     [Google Scholar]
  16. Lusis A. J., Becker R. R. 1973; The β-glucosidase system of the thermophilic fungus Chaetomium thermophile var.coprophile n. var. Biochimica et biophysica acta 329:5–16
     [Google Scholar]
  17. Miller G. L., Dean J., Blum R. 1960; A study of methods for preparing oligosaccharides from cellulose. Archives of Biochemistry and Biophysics 91:21–26
     [Google Scholar]
  18. Park J. T., Johnson M. S. 1949; Submicrodetermination of glucose. Journal of Biological Chemistry 181:149–151
     [Google Scholar]
  19. Reese E. T. 1977; Degradation of polymeric carbohydrates by microbial enzymes. Recent Advances in Phytochemistry 11:311–365
     [Google Scholar]
  20. Roth W. W., Srinivasan V. R. 1978; Affinity chromatographic purification of β-glucosidase of Candida guilliermondii. Preparative Biochemistry 8:57–71
     [Google Scholar]
  21. Rudick M. J., Elbein A. D. 1973; Glycoprotein enzymes secreted by Aspergillus fumigatus. Purification and properties of β-glucosidase. Journal of Biological Chemistry 248:6506–6513
     [Google Scholar]
  22. Sano K., Amemura A., Harada T. 1975; Purification and properties of a β-1,6-glucosidase from Flavobacterium. Biochimica et biophysica acta 377:410–420
     [Google Scholar]
  23. Shinmyo A., Garcia Martinez D. V., Demain A. L. 1979; Studies on the extracellular cellulolytic enzyme complex produced by Clostridium thermocellum. Journal of Applied Biochemistry 1:202–209
     [Google Scholar]
  24. Sternberg D., Viyayakumar P., Reese E. 1976; β-Glucosidase microbial production and effect on enzymatic hydrolysis of cellulose. Canadian Journal of Microbiology 23:139–147
     [Google Scholar]
  25. Trevelyan W. E., Procter D. P., Harrison J. S. 1950; Detection of sugars on paper chromatograms. Nature; London: 166444–445
     [Google Scholar]
  26. Umezurike G. M. 1971; The purification and properties of extracellular β-glucosidase from Botryodiplodia theobromae Pat. Biochimica et biophysica acta 227:419–428
     [Google Scholar]
  27. Weber K., Osborn M. 1969; The reliability of molecular weight determinations by dodecyl sulphate polyacrylamide gel electrophoresis. Journal of Biological Chemistry 244:4406–4412
     [Google Scholar]
/content/journal/micro/10.1099/00221287-128-3-569
Loading
Properties of β-Glucosidase Purified from Clostridium thermocellum
Microbiology 128, 569 (1982); https://doi.org/10.1099/00221287-128-3-569
/content/journal/micro/10.1099/00221287-128-3-569
/content/journal/micro/10.1099/00221287-128-3-569
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error