1887

Abstract

1,3--Glucanase I is one of the three 1,3--glucanase enzymes which are developmentally regulated in . On incubation of the fungus in derepressing medium (limited in glucose), 1,3--glucanases II and III are produced first, followed by 1,3--glucanase I. We have purified the latter enzyme 290-fold from cell-free extracts of derepressed mycelium. The purification involved Sephadex G-100 chromatography, adsorption to DEAE-Sephadex, concanavalin A-Sepharose and preparative isoelectric focusing. The purified enzyme appeared homogeneous on SDS-PAGE. The apparent molecular weights estimated by Sephadex G-100 filtration and SDS-PAGE were 65000 and 68000 respectively. The enzyme appears to be an acidic glycoprotein (pI 4·7) with an endo-splitting mode of action, clearly different from 1,3--glucanases II and III. The on a standard, slightly branched 1,3--glucan (laminarin) was 0·04 mg ml, at least ten times lower than the average m for -glucanases. It showed almost no activity on heavily branched substrates (yeast wall glucan). An evaluation of the physicochemical properties and other characteristics suggests a metabolic role for this endo-1,3--glucanase.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-128-9-2047
1982-09-01
2024-05-04
Loading full text...

Full text loading...

/deliver/fulltext/micro/128/9/mic-128-9-2047.html?itemId=/content/journal/micro/10.1099/00221287-128-9-2047&mimeType=html&fmt=ahah

References

  1. Abd-El-Al A., Phaff H. J. 1969; Purification and properties of endo-β-glucanases in the yeast Hanseniaspora valbyensis. Canadian Journal of Microbiology 15:697–701
    [Google Scholar]
  2. Bartnicki-Garcia S. 1973; Fundamental aspects of hyphal morphogenesis. Symposia of the Society for General Microbiology 23:245–267
    [Google Scholar]
  3. Chesters C. G. C., Bull A. T. 1963a; The enzymic degradation of laminarin. 1. The distribution of laminarinase among micro-organisms. Bio-chemical Journal 86:28–31
    [Google Scholar]
  4. Chesters C. G. C., Bull A. T. 1963b; The enzymic degradation of laminarin. 2. The multicomponent nature of fungal laminarinases. Biochemical Journal 86:31–38
    [Google Scholar]
  5. Clarke A. E., Stone B. A. 1965; β-Glucan hydrolase from Aspergillus niger. Isolation of a 1,4-β-glucan hydrolase and some properties of the 1,3-β-glucan hydrolase components. Biochemical Journal 96:793–801
    [Google Scholar]
  6. Davis B. J. 1964; Disc electrophoresis II. Method and application to human serum proteins. Annals of the New York Academy of Sciences 122:407–427
    [Google Scholar]
  7. Friebe B., Holldorf A. W. 1975; Control of ex-tracellular l,3-β-glucanase activity in a basidiomycete species. Journal of Bacteriology 122:818–825
    [Google Scholar]
  8. Mahadevan P. R., Mahadkar V. R. 1970; Role of enzyme in growth and morphology of Neurospora crassa: cell wall-bound enzymes and their possible role in branching. Journal of Bacteriology 101:941–947
    [Google Scholar]
  9. Northcote D. H., Horne R. D. 1952; The chemical composition and structure of the yeast cell wall. Biochemical Journal 51:232–236
    [Google Scholar]
  10. Notario V., Villa T. G., Villanueva J. R. 1976; Purification of an exo-β-glucanase from cell-free extracts of Candida utilis. Biochemical Journal 159:555–562
    [Google Scholar]
  11. Reese E. T., Parrish F. W., Mandels M. 1962; 1,6-β-Glucanases in fungi. Canadian Journal of Microbiology 8:327–334
    [Google Scholar]
  12. Rey F., Acha I. G., Nombela C. 1979; The regulation of β-glucanase synthesis in fungi and yeasts. Journal of General Microbiology 110:83–89
    [Google Scholar]
  13. Santos T., Villanueva J. R., Nombela C. 1978a; Regulation of l,3-β-glucanase synthesis in Penicillium italicum. Journal of Bacteriology 133:542–548
    [Google Scholar]
  14. Santos T., SÁnchez M., Villanueva J. R., Nombela C. 1978; . Regulation of the 1,3-β-glucanase system of Penicillium italicum: glucose repression of the various enzymes. Journal of Bacteriology 133:465–471
    [Google Scholar]
  15. Santos T., SÁnchez M., Villanueva J. R., Nombela C. 1979a; Derepression of 1,3-β-glucanases in Penicillium italicum: localization of the various enzymes and correlation with cell wall glucan mobilization and autolysis. Journal of Bacteriology 137:6–12
    [Google Scholar]
  16. Santos T., Rey F., Conde J., Villanueva J. R., Nombela C. 1979b; Saccharomyces cerevisiae mutant defective in exo-l,3-β-glucanase production. Journal of Bacteriology 139:333–338
    [Google Scholar]
  17. Sentandreu R., Elorza M. V., Villanueva J. R. 1975; Synthesis of yeast wall glucan. Journal of General Microbiology 90:13–20
    [Google Scholar]
  18. Trevelyan W. E., Procter W. E., Harrison J. S. 1950; Detection of sugars on paper chromato-grams. Nature; London: 166444–445
    [Google Scholar]
  19. Villanueva J. R., Notario V., Santos T., Villa T. G. 1976; β-Glucanases in nature. In Microbial and Plant Protoplasts pp. 323–355 Peberdy J. F., Rose A. H., Rogers H. J., Cocking E. C. Edited by London: Academic Press;
    [Google Scholar]
  20. Weber K., Osborn M. 1969; The reliability of molecular weight determination by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Journal of Biological Chemistry 244:4406–4412
    [Google Scholar]
  21. Wessels J. G. H. 1966; Control of cell-wall glucan degradation during development in Schizophyllum commune. Antonie van Leeuwenhoek 32:341–355
    [Google Scholar]
  22. Wessels J. G. H., Koltin Y. 1972; R-glucanase activity and susceptibility of hyphal walls to degradation in mutants of Schizophyllum with disrupted nuclear migration. Journal of General Microbiology 71:471–475
    [Google Scholar]
  23. Zacharius R. M., Zell T. E., Morrison J. H., Woodlock J. J. 1969; Glycoprotein staining following electrophoresis on acrylamide gels. Analytical Biochemistry 30:148–152
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-128-9-2047
Loading
/content/journal/micro/10.1099/00221287-128-9-2047
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