1887

Abstract

A chitosanase was purified from the intercellular fluid extract of chemically stressed barley () leaves. Purification was achieved by preparative PAGE involving separation under native conditions at pH 4.3 (Reisfeld system) followed by denaturing PAGE in the presence of SDS. One basic chitosanase with a molecular mass of 19 kDa could hydrolyse chitosan and glycol chitosan of crustacean origin in addition to fungal chitosan isolated after treatment of cells with sodium hydroxide followed by extraction in acetic acid. This enzyme did not exhibit activity towards -1,4-polymers such as chitin, cellulose or peptidoglycan (from ) after SDS-PAGE. The chitosanase was conjugated to colloidal gold at pH 9·5 and applied to fungal tissue sections or to isolated glycol chitosan, chitosan, cellulose or chitin. The chitosanase-gold complex reacted intensely with glycol chitosan and chitosan but did not bind to chitin or cellulose. The enzyme-gold complex also labelled chitosan isolated from . However, it did not react with cells in tissue sections. Labelling with the chitosanase-gold complex was easily detected over cell walls of f. sp. spores and hyphae as well as over cell walls of and . No binding was observed with .

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-137-8-2007
1991-08-01
2021-08-03
Loading full text...

Full text loading...

/deliver/fulltext/micro/137/8/mic-137-8-2007.html?itemId=/content/journal/micro/10.1099/00221287-137-8-2007&mimeType=html&fmt=ahah

References

  1. Araki Y., ITO E. 1988; Chitin deacetylase. Methods in Enzymology 161:510–514
    [Google Scholar]
  2. Asselin A., Grenier J., Côté F. 1985; Light-influenced extracellular accumulation of b (pathogenesis-related) proteins in Nicotiana green tissue induced by various chemicals or prolonged floating on water. Canadian Journal of Botany 63:1276–1283
    [Google Scholar]
  3. Audy P., Trudel J., Asselin A. 1988; Purification and characterization of a lysozyme from wheat germ. Plant Science 58:43–50
    [Google Scholar]
  4. Audy P., Grenier J., Asselin A. 1989; Lysozyme activity in animal extracts after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Comparative Biochemistry and Physiology 92B:523–527
    [Google Scholar]
  5. Bartnicki-Garcia S. 1968; Cell wall chemistry, morphogenesis, and taxonomy of fungi. Annual Review of Microbiology 22:87–108
    [Google Scholar]
  6. Benhamou N. 1988; Preparation and application of lectin-gold complexes. In Colloidal Gold Techniques and Applications95–143 Hayat M. A. New York: Academic Press;
    [Google Scholar]
  7. Benhamou N. 1989; Cytochemical localization of β-(l→4)-d-glucans in plant and fungal cells using an exoglucanase-gold complex. In Electron Microscopy Reviews : Subcellular and Biomolecular Structure123–138 Harris J. R. London: Pergamon Press;
    [Google Scholar]
  8. Benhamou N., Asselin A. 1989; Attempted localization of a substrate for chitinases in plant cells reveals abundant N-acetyl-d-glucosamine residues in secondary walls. Biology of the Cell 61:341–350
    [Google Scholar]
  9. Benhamou N., Chamberland H., Ouellette G. B., Pauze F. J. 1987; Ultrastructural localization of β-(l→4)-d-glucans in two pathogenic fungi and in their host tissues by means of an exoglucanase-gold complex. Canadian Journal of Microbiology 33:405–417
    [Google Scholar]
  10. Benhamou N., Chamberland H., Noel S., Ouellette G. B. 1990; Ultrastructural localization of β-l,4-glucan-containing molecules in the cell walls of some fungi: a comparative study between spore and mycelium. Canadian Journal of Microbiology 36:149–158
    [Google Scholar]
  11. Briza P., Ellinger A., Wrikler G., Breitenbach M. 1988; Chemical composition of the yeast ascospore wall. The second outer layer consists of chitosan. Journal of Biological Chemistry 263:11569–11574
    [Google Scholar]
  12. Côté F., El Ouakfaoui S., Asselin A. 1991; Detection of β-glucanase activity on various β-1,3- and β-1,4-glucans after native and denaturing polyacrylamide gel electrophoresis. Electrophoresis 12:69–74
    [Google Scholar]
  13. Datema R., Van den Ende H., Wessels J. G. H. 1977a; The hyphal wall of Mucor mucedo 1. Polyanionic polymers. European Journal of Biochemistry 80:611–619
    [Google Scholar]
  14. Datema R., Wessels J. G. H., Van den Ende H. 1977b; The hyphal wall of Mucor mucedo. 2 Hexosamine-containing polymers. European Journal of Biochemistry 80:621–626
    [Google Scholar]
  15. Fenton D. M., Eveleigh D. E. 1981; Purification and mode of action of a chitosanase from Penicillium islandicum . Journal of General Microbiology 126:151–165
    [Google Scholar]
  16. Frens G. 1973; Controlled nucleation for regulation of the particle size in monodisperse gold suspensions. Nature Physical Science 241:20–22
    [Google Scholar]
  17. Grenier J., Asselin A. 1990; Some pathogenesis-related proteins are chitosanases with lytic activity against fungal spores. Molecular Plant-Microbe Interactions 3:401–407
    [Google Scholar]
  18. Hadwiger L. A., Line R. F. 1981; Hexosamine accumulations are associated with the terminated growth of Puccinia striiformis on wheat isolines. Physiological Plant Pathology 19:249–255
    [Google Scholar]
  19. Hadwiger L. A., Beckman J. M., Adams M. J. 1981; Localization of fungal components in the pea-Fusarium interaction detected immunochemically with anti-chitosan and anti-fungal cell wall antisera. Plant Physiology 67:170–175
    [Google Scholar]
  20. Kendra D. F., Christian D., Hadwiger L. A. 1989; Chitosan oligomers from Fusarium solani/pea interactions, chitinase/β-glucan-ase digestion of sporelings and from fungal wall chitin actively inhibit fungal growth and enhance disease resistance. Physiological and Molecular Plant Pathology 35:215–230
    [Google Scholar]
  21. Letourneau D. R., Deven J. M., Manocha M. S. 1976; Structure and composition of the cell wall of Choanephora cucurbitarum . Canadian Journal of Microbiology 22:486–494
    [Google Scholar]
  22. McCormick J. J., Blomquist J. C., Rusch H. P. 1970; Isolation and characterization of a galactosamine wall from spores and spherules of Physarum polycephalum . Journal of Bacteriology 104:1119–1125
    [Google Scholar]
  23. McGahren W. J., Perkinson G. A., Growich J. A., Leese R. A., Ellestad G. A. 1984; Chitosan by fermentation. Process Biochemistry 8:88–90
    [Google Scholar]
  24. Mol P. C., Wessels J. G. H. 1987; Linkages between glucosaminoglycan and glucan determine alkali-insolubility in walls of Saccharomyces cerevisiae . FEMS Microbiology Letters 41:95–99
    [Google Scholar]
  25. Monaghan R. L., Eveleigh D. E., Tewarin R. P., Reese E. T. 1973; Chitosanase, a novel enzyme. Nature New Biology 245:78–80
    [Google Scholar]
  26. Novaes-Ledieu M., Garcia Mendoza D. 1981; The cell walls of Agaricus bisporus and Agaricus campestris fruiting body hyphae. Canadian Journal of Microbiology 27:779–787
    [Google Scholar]
  27. Parent J.-G., Asselin A. 1984; Detection of pathogenesis-related proteins (PR or b) and of other proteins in the intercellular fluid of hypersensitive plants infected with tobacco mosaic virus. Canadian Journal of Botany 62:564–569
    [Google Scholar]
  28. Pelletier A., Syguch J. 1990; Purification and characterization of 3 chitosanase activities from Bacillus megaterium PI. Applied and Environmental Microbiology 56:844–848
    [Google Scholar]
  29. Qureshi A. A., Tage O. T. 1970; Observations on chlamydospore production by Fusarium in a two-salt solution. Canadian Journal of Microbiology 16:29–32
    [Google Scholar]
  30. Reyes F., Lahoz R., Martinez M. J., Alfonso C. 1985; Chitosanases in the autolysis of Mucor rouxii . Mycopathologia 89:181–187
    [Google Scholar]
  31. Sandford P. A., Hutchings G. P. 1987; Chitosan - a natural, cationic biopolymer: commercial applications. In Industrial Polysac-charides: Genetic Engineering, Structure I Property Relations and Applications363–375 Yalpani M. Amsterdam: Elsevier;
    [Google Scholar]
  32. Siegrist J., Kauss H. 1990; Chitin deacetylase in cucumber leaves infected by Colletotrichum lagenarium . Physiological and Molecular Plant Pathology 36:267–275
    [Google Scholar]
  33. Trudel J., Asselin A. 1989; Detection of chitinase activity after polyacrylamide gel electrophoresis. Analytical Biochemistry 178:362–366
    [Google Scholar]
  34. Trudel J., Asselin A. 1990; Detection of chitin deacetylase activity after polyacrylamide gel electrophoresis. Analytical Biochemistry 189:249–253
    [Google Scholar]
  35. Trudel J., Audy P., Asselin A. 1989; Electrophoretic forms of chitinase activity in Xanthi-nc tobacco, healthy and infected with tobacco mosaic virus. Molecular Plant-Microbe Interactions 2:315–324
    [Google Scholar]
  36. Wessels J. G. H., Sietsma J. H. 1981; Fungal cell walls: a survey. In Plant Carbohydrates II352–394 Tanner W., Loewus F. A. New York: Springer-Verlag;
    [Google Scholar]
  37. White S. A., Farina P. R., Fulton I. 1979; Production and isolation of chitosan from Mucor rouxii . Applied and Environmental Microbiology 38:323–328
    [Google Scholar]
  38. Yokoyama T., Murakami E., Hasegawa K., Tukada S., Takagi H., Kadowaki K., Oishi K. 1989; Microbial production of poly hexosamine. In Chitin and Chitosan333–343 Skjak-Braek G., Anthonsen T., Sandford P. London: Elsevier Applied Science;
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-137-8-2007
Loading
/content/journal/micro/10.1099/00221287-137-8-2007
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