1887

Abstract

Chitinase activities were detected in the supernatant and microsomal fractions from homogenized mycelium of surface-grown cultures of . Most properties of the two preparations were very similar, e.g. pH optima were 5·65 and 5·55, respectively, and values were 16·7 and 19·2 mg chitin ml, respectively. Neither activity had a cationic requirement, but both were inhibited by CuSome properties differed. On freezing, the supernatant activity remained constant over 4 d and then gradually decreased, whereas microsomal activity increased, suggesting that endogenous activation was occurring. Prolonged incubations at 30 °C also allowed endogenous activation of microsomal activity, which could be prevented by incubation with phenylmethanesulphonyl fluoride. This enzyme activity could also be activated by treatment of the microsomes with proteases such as trypsin. Inactivation occurred after treatment with phospholipases, suggesting that microsomal chitinase requires phospholipid-enzyme interactions for activity. Thus the microsomal activity behaved as a membrane-bound zymogen.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-130-6-1359
1984-06-01
2021-10-20
Loading full text...

Full text loading...

/deliver/fulltext/micro/130/6/mic-130-6-1359.html?itemId=/content/journal/micro/10.1099/00221287-130-6-1359&mimeType=html&fmt=ahah

References

  1. Achstetter T., Ehmann C., Wolf D. H. 1980; New proteolytic enzymes in yeast. Archives of Biochemistry and Biophysics 207:445–454
    [Google Scholar]
  2. Algranati I. D. 1963; Determination of initial rates in enzymic non-linear progress reactions. Biochimica et biophysica acta 73:152–155
    [Google Scholar]
  3. Araki Y., Ito E. 1975; A pathway of chitosan formation in Mucor rouxii. European Journal of Biochemistry 55:71–78
    [Google Scholar]
  4. Aribisala O. A., Gooday G. W. 1978; Properties of chitinase from Vibrio alginolyticus as assayed with the chromogenic substrate 3,4-dinitrophenol-tetra-N-acety-β-D-chitotetraoside. Biochemical Society Transactions 6:568–569
    [Google Scholar]
  5. Bartnicki-Garcia S. 1973; Fundamental aspects of hyphal morphogenesis. Symposia of the Society for General Microbiology 23:245–267
    [Google Scholar]
  6. Bartnicki-Garcia S., Davis L. L. 1983; Chitosan synthesis in Mucor rouxii: mechanism and regulation. Abstracts of the 3rd International Mycological Congress p. 15 Tokyo: Science Council of Japan;
    [Google Scholar]
  7. Berkeley R. W. C. 1979; Chitin, chitosan and their degradative enzymes. In Microbial Polysaccharides and Poly saccharoses pp. 205–236 Edited by Berkeley R. C. W, Gooday G. W., Ellwood D. C. London: Academic Press;
    [Google Scholar]
  8. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254
    [Google Scholar]
  9. Cabib E., Duran A., Bowers B. 1979; Localized activation of chitin synthetase in the initiation of yeast septum formation. In Fungal Walls and Hyphal Growth pp. 189–201 Edited by Burnett J. H., Trinci A. P. J. Cambridge: Cambridge University Press;
    [Google Scholar]
  10. Elango N., Correa J. U., Cabib E. 1982; Secretory character of yeast chitinase. Journal of Biological Chemistry 257:1398–1400
    [Google Scholar]
  11. Gooday G. W., Trinci A. P. J. 1980; Wall structure and biosynthesis in fungi. Symposia of the Society of General Microbiology 30:207–251
    [Google Scholar]
  12. Hardy J. C., Gooday G. W. 1983; Stability and zymogenic nature of chitin synthase from Candida albicans. Current Microbiology 9:51–54
    [Google Scholar]
  13. Iten W., Matile P. 1970; Role of chitinase and other lysosomal enzymes of Coprinus lagopus in the autolysis of fruiting bodies. Journal of General Microbiology 61:301–309
    [Google Scholar]
  14. Lopez-Romero E., Ruiz-Herrera J., Bartnicki-Garcia S. 1982; The inhibitory protein of chitin synthase from Mucor rouxii is a chitinase. Biochimica et biophysica acta 702:233–236
    [Google Scholar]
  15. Molano J., Duran A., Cabib E. 1977; A rapid and sensitive assay for chitinase using tritiated chitin. Analytical Biochemistry 83:648–656
    [Google Scholar]
  16. Muzzarelli R. A. A, Tanfani F., Scarpini G., Laterza G. 1980; The degree of acetylation of chitins by gas chromatography and infrared spec-troscopy. Journal of Biochemical and Biophysical Methods 2:299–306
    [Google Scholar]
  17. North M. J. 1982; Comparative biochemistry of the proteinases of eukaryotic microorganisms. Microbiological Reviews 46:308–340
    [Google Scholar]
  18. Polachek I., Rosenberger R. F. 1978; Distri-bution of autolysins in hyphae of Aspergillus nidulans: evidence for a lipid-mediated attachment to hyphal walls. Journal of Bacteriology 135:741–747
    [Google Scholar]
  19. Rosenberger R. F. 1979; Endogenous lytic enzymes and wall metabolism. In Fungal Walls and Hyphal Growth pp. 265–277 Edited by Burnett J. H., Trinci A. P. J. Cambridge: Cambridge University Press;
    [Google Scholar]
  20. Ulane R. E., Cabib E. 1976; The activating system of chitin synthetase from Saccharomyces cerevisiae. Journal of Biological Chemistry 251:3367–3374
    [Google Scholar]
  21. Wilson J. E. 1978; Ambiquitous enzymes: variation in intracellular distribution as a regulatory mechanism. Trends in Biochemical Sciences 3:124–125
    [Google Scholar]
  22. Zarain-Herzberg A., Arroyo-Begovich A. 1983; Chitinolytic activity from Neurospora crassa. Journal of General Microbiology 129:3319–3326
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-130-6-1359
Loading
/content/journal/micro/10.1099/00221287-130-6-1359
Loading

Data & Media loading...

Most cited this month 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