Properties of the Endopolygalacturonase Secreted by Rhizopus stolonifer Free

Abstract

The endopolygalacturonase secreted by Ehr. ex Fr. during infection of strawberries of the cultivar ‘ Cambridge Favourite’ was extracted with a solution of sodium chloride and partially purified by gel filtration and ion-exchange chromatography. Partial purification removed 99·8 % of the contaminating uronide materials, but the enzyme yield was reduced to 56 % and the specific activity was increased only 2·5 times. The partially purified enzyme exhibited maximum stability at pH 4·0 to 6·0 and optimal activity at pH 4·6 to 4·8.A linear thermal-inactivation pattern between 30 and 50 °C was demonstrated and complete and irreversible inactivation was achieved by heating to 60 °C for 20 min. Enzyme activity was inhibited by 20 or less by a range of enzyme inhibitors and cations, with 87 % inhibition occurring in the presence of 10 -Hg. The molecular weight of the enzyme was calculated as 60000, and the sedimentation and diffusion data suggested that the enzyme molecule has a high degree of asymmetry.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-80-2-401
1974-02-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/80/2/mic-80-2-401.html?itemId=/content/journal/micro/10.1099/00221287-80-2-401&mimeType=html&fmt=ahah

References

  1. Andrews P. 1964; Estimation of the molecular weight of proteins by Sephadex gel filtration. Biochemical Journal 91:222–233
    [Google Scholar]
  2. Barash I., Eyal Z. 1970; Properties of a polygalacturonase produced by Geotrichum candidum . Phytopathology 60:27–30
    [Google Scholar]
  3. Bateman D. F. 1968; The enzymatic maceration of plant tissue. Netherlands Journal of Plant Pathology 74 (S1):67–80
    [Google Scholar]
  4. Bateman D. F., Millar R. L. 1966; Pectic enzymes in tissue degradation. Annual Review of Phytopathology 4:119–146
    [Google Scholar]
  5. Bitter T., Muir H. M. 1962; A modified carbazoleuronic acid reaction. Analytical Biochemistry 4:330–334
    [Google Scholar]
  6. Cappellini R. A. 1966; Growth and polygalacturonase production by Rhizopus stolonifer . Phytopathology 56:734–737
    [Google Scholar]
  7. Dakin J. C., Tampion J. 1968; Specific fungi as the causative agents of the disintegration of sulphited strawberries. Journal of Food Technology 3:39–48
    [Google Scholar]
  8. Fielding A. H., Byrde R. J. W. 1969; The partial purification and properties of endopolygalacturonase and α-l-arabinofuranosidase secreted by Sclerotinia fructigena . Journal of General Microbiology 58:73–84
    [Google Scholar]
  9. Jansen E. F., MacDonnell L. R. 1945; Influence of methoxyl content of pectic substances on the action of polygalacturonase. Archives of Biochemistry 8:97–112
    [Google Scholar]
  10. Kumari L. H., Sirsi I. 1971; Purification and properties of endopolygalacturonase from Ganoderma lucidum . Journal of General Microbiology 65:285–290
    [Google Scholar]
  11. Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. 1951; Protein measurements with Folin phenol reagent. Journal of Biological Chemistry 193:265–275
    [Google Scholar]
  12. McComb E. A., McCready R. M. 1952; Colorimetric determination of pectic substances. Analytical Chemistry 24:1630–1632
    [Google Scholar]
  13. Nelson N. 1944; A photometric adaptation of the Somogyi method for the determination of glucose. Journal of Biological Chemistry 153:373–380
    [Google Scholar]
  14. Schachman H. K. 1957; Ultracentrifugation, diffusion and viscometry. In Methods in Enzymology 4 pp. 32–103 Colwick S. P., Kaplan N. O. Edited by New York: Academic Press;
    [Google Scholar]
  15. Selby K. 1961; The degradation of cotton cellulose by the extracellular cellulase of Myrothecium verrucaria . Biochemical Journal 79:562–566
    [Google Scholar]
  16. Srivastava D. N., Echandi E., Walker J. C. 1959; Pectolytic and cellulolytic enzymes produced by Rhizopus stolonifer . Phytopathology 49:145–148
    [Google Scholar]
  17. Swinburne T. R., Corden M. E. 1967; Dissociation and recombination of a polygalacturonase complex during ion-exchange chromatography. Nature; London: 213286–287
    [Google Scholar]
  18. Trescott A. S. 1971 A study of the enzymes secreted by Rhizopusstolonifer Ehr. ex Fr. which are responsible for the disintegration of sulphited strawberries (Fragaria ‘Cambridge Favourite’) Ph.D. Thesis London University:
    [Google Scholar]
  19. Wood R. K. S. 1955; Pectic enzymes secreted by pathogens and their role in plant infection. In Mechanisms of Microbial Pathogenicity pp. 263–293 Howie J. W., O’Hea A. J. Edited by Cambridge University Press;
    [Google Scholar]
  20. Wood R. K. S. 1960; Pectic and cellulolytic enzymes in plant disease. Annual Review of Plant Physiology 11:299–322
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-80-2-401
Loading
/content/journal/micro/10.1099/00221287-80-2-401
Loading

Data & Media loading...

Most cited Most Cited RSS feed