1887

Abstract

Chitin synthase activity was detected in actively growing mycelium of after mechanical disruption of the cells. Chitin formation in fungal extracts at 32 °C was linear with respect to time for at least 60 min, and with respect to protein concentration up to 750 μg ml. The optimum pH for enzyme activity was 8·5 using 10 m-Tris/HCl buffer. Mg was necessary for maximum activity and 10 m-MgC1 was routinely used during the assays. The apparent for the substrate UDP-GlcNAc was 2 m. Polyoxin D was a competitive inhibitor of chitin synthesis with an apparent of 4 μ. Following treatment with trypsin (12·5 μg ml), the chitin synthase activity of the fungal extract increased by six-fold, indicating that most of the chitin synthase activity was zymogenic. The reaction product was insoluble in 1 -KOH or 1 -acetic acid, but it was solubilized by heating in 6 -HCl at 120 °C for 2.5 h and was hydrolysed by chitinase into diacetylchitobiose.

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/content/journal/micro/10.1099/00221287-135-2-279
1989-02-01
2024-04-23
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References

  1. Cabib E. 1981; Chitin: structure, metabolism and regulation of biosynthesis. In Encyclopedia of Plant Physiology New Series 13B Plant Carbohydrates II, pp. 395–415 Tanner W., Loewus F.A. Edited by Berlin: Springer Verlag;
    [Google Scholar]
  2. Cabib E., Duran A., Bowers B. 1979; Localized action of chitin synthase in the initiation of yeast septum formation. In Fungal Walls and Hyphal Growth pp. 189–201 Burnett J.H., Trinci A.P.J. Edited by Cambridge: Cambridge University Press;
    [Google Scholar]
  3. Endo A., Kakiki K., Misato T. 1970; Mechanisms of action of antifungal agent polyoxin D. Journal of Bacteriology 105:161–164
    [Google Scholar]
  4. Gooday G.W. 1978; The enzymology of hyphal growth. In Developmental Mycology 3 The Filamentous Fungi pp. 51–77 Smith J.E., Berry D.R. Edited by London: Edward Arnold;
    [Google Scholar]
  5. Gooday G.W., Derousset-Hall A. 1975; Properties of chitin synthetase from Coprinus ciner-eus . Journal of General Microbiology 89:137–145
    [Google Scholar]
  6. Gooday G.W., Trinci A.P.J. 1980; Wall structure and biosynthesis in fungi. In The Eucaryotic Microbial Cell pp. 207–251 Gooday G.W., Lloyd D., Trinci A.P.J. Edited by Cambridge: Cambridge University Press;
    [Google Scholar]
  7. Hardy J.C., Gooday G.W. 1978; Chitin biosynthesis by Candida albicans . Proceedings of the Society for General Microbiology 5:106–107
    [Google Scholar]
  8. Lowe S.E., Theodorou M.K., Trinci A.P.J. 1985; Growth of anaerobic rumen fungi on defined and semi-defined media lacking rumen fluid. Journal of General Microbiology 131:2225–2229
    [Google Scholar]
  9. Orlean P. 1987; Two chitin synthases in Saccharo-myces cerevisiae . Journal of Biological Chemistry 262:5732–5739
    [Google Scholar]
  10. Orpin C.G. 1977; The occurrence of chitin in the cell walls of rumen organisms Neocallimastix frontalis, Piromonas communis and Sphaeromonas communis . Journal of General Microbiology 99:215–218
    [Google Scholar]
  11. Raeven M.B.J.M. 1981; Biosynthesis of chitin in fungi. PhD thesis University of Groningen; The Netherlands:
    [Google Scholar]
  12. De Rousset-Hall A., Gooday G.W. 1975; A kinetic study of a solubilized chitin synthetase preparation from Coprinus cinereus . Journal of General Microbiology 89:146–159
    [Google Scholar]
  13. Sburlati A., Cabib E. 1986; Chitin synthetase,2 a presumptive participant in septum formation in Saccharomyces cerevisiae . Journal of Biological Chemistry 261:15147–15152
    [Google Scholar]
  14. Vermeulen C.A., Raeven M.B.J.M., Wessels J.G.H. 1979; Localization of chitin synthase activity in subcellular fractions of Schizophyllum commune protoplasts. Journal of General Microbiology 114:87–97
    [Google Scholar]
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