1887

Abstract

An exo-1,3 -glucanase was purified from blastoconidia of 1001. The purified enzyme appeared as a single protein band by PAGE, and split into two subunits ( approximately 63000 and 44000) when analysed by SDS-PAGE. The pI of the enzyme was 4 and a of 1·7 mg ml was estimated for laminarin as substrate. Despite its very reduced activity on the synthetic substrate -nitrophenyl --glucoside, exo-1,3--glucanase hydrolysed 1,3--glucan by an exo-splitting mechanism and was inhibited by glucono--lactone and by Hg and Ag cations. The active exo-glucanase was mainly located in the periplasm, but it was also present inside the cytoplasmic membrane in small amounts and was secreted into the culture medium. The electrophoretic mobility of the enzyme from all three locations was the same.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-135-2-309
1989-02-01
2024-04-25
Loading full text...

Full text loading...

/deliver/fulltext/micro/135/2/mic-135-2-309.html?itemId=/content/journal/micro/10.1099/00221287-135-2-309&mimeType=html&fmt=ahah

References

  1. Bull A.T. 1967; The enzymatic degradation of β-glucans. International Biodeterioration Bulletin 3:3–6
    [Google Scholar]
  2. Cenamor R., Molina M., Galdona J., Sanchez M., Nombela C. 1987; Production and secretion of Saccharomyces cerevisiae β -glucanases: differences between protoplast and periplasmic enzymes. Journal of General Microbiology 133:619–628
    [Google Scholar]
  3. Goldstein I.J., Hay G.W., Lewis B.A., Smith F. 1965; Controlled degradation of polysaccharides by periodate oxidation reduction and hydrolysis. Methods in Carbohydrate Chemistry 5:361–369
    [Google Scholar]
  4. Gopal P.K., Shepherd M.G., Sullivan P.A. 1984; Analysis of wall glucan from yeast, hyphal and germ tube forming cells ofCandida albicans . Journal of General Microbiology 130:3295–3301
    [Google Scholar]
  5. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature London: 227:680–685
    [Google Scholar]
  6. Merril C.R., Goldman D., VanKeuren M.L. 1982; Simplified silver protein detection and image enhancement methods in polyacrylamide gels. Electrophoresis 3 :17–21
    [Google Scholar]
  7. Molina M., Cenamor R., Nombela C. 1987; Exo-1,3-β-glucanase activity inCandida albicans: effect of the yeast-to-mycelium transition. Journal of General Microbiology 133:609–617
    [Google Scholar]
  8. Notario V. 1982; β-Glucanases fromCandida albicans: purification, characterization and the nature of their attachment to the cell wall components. Journal of General Microbiology 128:747–759
    [Google Scholar]
  9. Odds F.C. 1985; Morphogenesis inCandida albicans . CRC Critical Reviews in Microbiology 12:45–80
    [Google Scholar]
  10. Ornstein L., Davis B.J. 1964; Disk electrophoresis. I. Background and theory. Annals of the New York Academy of Sciences 121:321–349
    [Google Scholar]
  11. Polacheck I., Melamed M., Bercovier H., Salkin I.F. 1987; β-Glucosidase in Candida albicans and its application to yeast identification. Journal of Clinical Microbiology 25:907–910
    [Google Scholar]
  12. Ram S.P., Romana L.K., Shepherd M.G., Sullivan P.A. 1984; Exo-l,3-β-glucanase, autoly-sin and trehalase activities during growth and germ tube formation in Candida albicans . Journal of General Microbiology 130:1227–1236
    [Google Scholar]
  13. Sanchez A., Nebreda A. R., Villa T. G. 1982a; Self-associating properties of yeast exo-β-glucanase are dependent on the ionic strength. FEBS Letters 145:213–216
    [Google Scholar]
  14. Sanchez A., Villanueva A.R., Villa T.G. 1982b; Effect of tunicamycin on exo-l,3-β-glucanase synthesis and secretion by cells and protoplasts ofSaccharomyces cerevisiae . Journal of General Microbiology 128:3051–3060
    [Google Scholar]
  15. Sullivan P.A., Chiew Y.Y., Molloy C., Templeton M.D., Shepherd M.G. 1983; An analysis of the metabolism and cell wall composition ofCandida albicans during germ tube formation. Canadian Journal of Biochemistry 125:209–212
    [Google Scholar]
  16. Villa T.G., Notario V., Villanueva J.R. 1975; β-Glucanases of the yeast Pichia polymorpha . Archives of Microbiology 104:201–206
    [Google Scholar]
  17. Villa T.G., Notario V., Benitez T., VillaNueva J.R. 1976; Purification of an exo-1,3-β-glucanase from Candida utilis . Canadian Journal of Biochemistry 54:927–934
    [Google Scholar]
  18. Villa T.G., Lachance M.A., Phaff H.J. 1978; β-Glucanases of the yeast Kluyveromyces phaseolo-sporus: partial purification and characterization. Experimental Mycology 2:12–25
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-135-2-309
Loading
/content/journal/micro/10.1099/00221287-135-2-309
Loading

Data & Media loading...

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