1887

Abstract

SUMMARY: Protoplasts were produced from mycelium using Trichoderma lytic enzyme. The influence of KC1 and MgSOas stabilizer systems on the morphological variation of protoplasts produced during digestion and the pattern of release from hyphae were compared. The results suggest that protoplast release in the presence of KC1 followed a sequential fractionation of the hyphae with ‘early’ protoplasts originating from the tip regions and ‘late’ protoplasts from the distal regions. In MgSO-stabilized systems the hyphae were disrupted in a less ordered fashion. Between 12 and 16% of the mycelial protein was recovered in protoplast form using the systems described.

The level of chitin synthase (EC 2.4.1.16) and the capacity for trypsin-activation of the enzyme in protoplast fractions was investigated. In KC1-stabilized systems, ‘early’ (1 h) fractions possessed higher specific activities than later fractions. Activatable enzyme was low in the early fraction but was present at high levels in later fractions. It is suggested that these observations are consistent with a model relating active and activatable enzyme to hyphal growth.

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1978-03-01
2024-03-28
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References

  1. Archer D. B. 1977; Chitin biosynthesis in protoplasts and sub-cellular fractions of Aspergillus fumigatus. Biochemical Journal 164:653–658
    [Google Scholar]
  2. Bartnicki-Garcia S. 1973; Fundamental aspects of hyphal morphogenesis. Symposia of the Society for General Microbiology 23:245–268
    [Google Scholar]
  3. Bartnicki-Garcia S., Lippman E. 1966; Liberation of protoplasts from the mycelium of Phytophthora. Journal of General Microbiology 42:411–416
    [Google Scholar]
  4. Cabib E. 1975; Molecular aspects of yeast morphogenesis. Annual Review of Microbiology 29:191–214
    [Google Scholar]
  5. De Vries O. M. H., Wessels J. G. H. 1972; Release of protoplasts from Schizophyllum commune by a lytic enzyme preparation from Trichoderma viride. Journal of General Microbiology 73:13–22
    [Google Scholar]
  6. De Vries O. M. H., Wessels J. G. H. 1975; Chemical analysis of cell wall regeneration and reversion of protoplasts from Schizophyllum commune. Archives of Microbiology 102:209–218
    [Google Scholar]
  7. Fawcett P. A., Loder P. B., Duncan M. J., Beesley T. J., Abraham E. P. 1973; Formation and properties of protoplasts from antibiotic-producing strains of Penicillium chrysogenum and Cephalosporium acremonium. Journal of General Microbiology 79:293–309
    [Google Scholar]
  8. Gibson R. K., Peberdy J. F. 1972; Fine structure of protoplasts of Aspergillus nidulans. Journal of General Microbiology 72:529–538
    [Google Scholar]
  9. Grove S. N., Bracker C. E. 1970; Protoplasmic organization of hyphal tips among fungi: vesicles and spitzenkorper. Journal of Bacteriology 104:989–1009
    [Google Scholar]
  10. Katz D., Rosenberger R. F. 1971; Hyphal wall synthesis in Aspergillus nidulans: effect of protein synthesis inhibition and osmotic stabiliser shock on chitin insertion and morphogenesis. Journal of Bacteriology 108:184–190
    [Google Scholar]
  11. Lopez-Romero E., Ruiz-Herrera J. 1976; Synthesis of chitin by particulate preparations from Aspergillus flavus. Antonie van Leeuwenhoek 42:261–276
    [Google Scholar]
  12. Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. 1951; Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
    [Google Scholar]
  13. Mcmurrough I., Bartnicki-Garcia S. 1973; Inhibition and activation of chitin synthesis by Mucor rouxii cell extracts. Archives of Biochemistry and Biophysics 158:812–816
    [Google Scholar]
  14. Peberdy J. F. 1976; Isolation and properties of protoplasts from filamentous fungi. In Microbial and Plant Protoplasts pp. 39–50 Edited by Peberdy J. F., Rose A. H., Rogers H. J., Cocking E. C. London, New York: Academic Press;
    [Google Scholar]
  15. Peberdy J. F., Isaac S. 1976; An improved procedure for protoplast isolation from Aspergillus nidulans. Microbios Letters 3:7–9
    [Google Scholar]
  16. Rej R., Richards A. H. 1974; Interference by tris buffer in the estimation of protein by the Lowry procedure. Analytical Biochemistry 62:240–247
    [Google Scholar]
  17. Ruiz-Herrera J., Bartnicki-Garcia S. 1976; Proteolytic activation and inactivation of chitin synthetase from Mucor rouxii. Journal of General Microbiology 97:241–249
    [Google Scholar]
  18. Ryder N. S. 1977 Chitin synthase in Aspergillus nidulans Ph.D. thesis, University of Nottingham
    [Google Scholar]
  19. Ryder N. S., Peberdy J. F. 1977; Chitin synthase in Aspergillus nidulans: properties and proteolytic activation. Journal of General Microbiology 99:69–76
    [Google Scholar]
  20. Sternlicht E., Katz D., Rosenberger R. F. 1973; Subapical wall synthesis and wall thickening induced by cycloheximide in hyphae of Aspergillus nidulans. Journal of Bacteriology 114:812–823
    [Google Scholar]
  21. Vallejo C. G., Lugunas R. 1970; Interference by sulphydryl, disulphide reagents and potassium ions on protein determination by Lowry’s method. Analytical Biochemistry 36:207–212
    [Google Scholar]
  22. Villanueva J. R., Garcia Acha I. 1971; Production and use of fungal protoplasts. Methods in Microbiology 4:665–718
    [Google Scholar]
  23. Zalokar M. 1965; Integration of cellular metabolism. In The Fungi 1 pp. 377–428 Edited by Ainsworth G. C., Sussman A. S. New York, London: Academic Press;
    [Google Scholar]
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