1887

Abstract

Various chemical reagents known to extract spore coat protein were used to extract spore-lytic enzyme (SLE) from intact and germinated spores of Of the reagents tested, 7·2-urea plus 10% (v/v) mercaptoethanol, pH2·85, solubilized the most SLE activity per mg spores. The quantity of SLE extracted was dependent on the initial pH of the reagent, with a maximum between pH 2·7 and 3·0. Germinated spores yielded more SLE than non-germinated spores upon urea/mercaptoethanol extraction. SLE release during spore germination probably utilizes a trigger mechanism not satisfied by germination alone. Significant amounts of SLE were released during germination when spores were suspended in potassium chloride or a complex germinant mixture containing brain-heart infusion, yeast extract and chloramphenicol, but not during germination with sodium nitrite, which non-enzymically lysed the cortical peptidoglycan. Greater solubilization of SLE activity was obtained by urea/mercaptoethanol extraction of spores germinated with nitrite than of spores germinated with either potassium chloride or the complex germinant.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-126-1-37
1981-09-01
2024-12-07
Loading full text...

Full text loading...

/deliver/fulltext/micro/126/1/mic-126-1-37.html?itemId=/content/journal/micro/10.1099/00221287-126-1-37&mimeType=html&fmt=ahah

References

  1. Ando Y. 1979; Spore lytic enzyme released from Clostridium perfringens spores during germination. Journal of Bacteriology 140:59–64
    [Google Scholar]
  2. Aronson A. I., Fitz-James P. C. 1976; Structure and morphogenesis of the bacterial spore coat. Bacteriological Reviews 40:360–402
    [Google Scholar]
  3. 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]
  4. Brown W. C., Cuhel R. L. 1975; Surfacelocalized cortex-lytic enzyme in spores of Bacillus cereus T. Journal of General Microbiology 91:429–432
    [Google Scholar]
  5. Brown W. C., Cuhel R. L., Greer C. 1977; Isolation and properties of a surface-bound cortexlytic enzyme from spores of Bacillus cereus T. In Spore Research 1976 pp. 335–349 Barker A., Wolf L., Ellar D. J., Dring G. J., Gould G. W. Edited by London: Academic Press.;
    [Google Scholar]
  6. Brown W. C., Vellom D., Schnepf E., Greer C. 1978; Purification of a surface-bound hexosaminidase from spores of Bacillus cereus T. FEMS Microbiology Letters 3:247–251
    [Google Scholar]
  7. Cassier M., Ryter A. 1971; Sur un mutant de Clostridium perfringens donnant des spores sans tuniques a germination lysozyme-dépendante. Annales de l’Institut Pasteur 117:312–324
    [Google Scholar]
  8. Cleveland E. F., Gilvarg C. 1975; Selective degradation of peptidoglycan from Bacillus megaterium spores. In Spores VI pp. 458–464 Gerhardt P., Costilow R. N., Sadoff H. L. Edited by Washington, D.C.: American Society for Microbiology.;
    [Google Scholar]
  9. Duncan C. L., Labbe R. G., Reich R. R. 1972; Germination of heat- and alkali-altered spores of Clostridium perfringens type A by lysozyme and an initiation protein. Journal of Bacteriology 109:550–559
    [Google Scholar]
  10. Franceschini T. J., Labbe R. G. 1979; Sensitivity of chemically treated spores of Clostridium perfringens type A to an initiation protein. Microbios 25:85–91
    [Google Scholar]
  11. Gould G. W., Hitchins A. D. 1963; Sensitization of bacterial spores to lysozyme and to hydrogen peroxide with agents which rupture disulphide bonds. Journal of General Microbiology 33:413–423
    [Google Scholar]
  12. Gould G. W., Hitchins A. D., King W. L. 1966; Function and location of a ‘germination enzyme’ in spores of Bacillus cereus. Journal of General Microbiology 44:293–302
    [Google Scholar]
  13. Hashimoto T., Frieben W., Conti S. 1972; Kinetics of germination of heat-injured Bacillus cereus spores. In Spores V pp. 409–415 Halvorson H. O., Hanson R., Campbell L. L. Edited by Washington, D.C.: American Society for Microbiology.;
    [Google Scholar]
  14. Labbe R. G., Reich R. R., Duncan C. L. 1978; Alteration in ultrastructure and germination of Clostridium perfringens type A spores following extraction of spore coats. Canadian Journal of Microbiology 24:1526–1536
    [Google Scholar]
  15. Powell J. F., Strange R. E. 1953; Biochemical changes occurring during germination of bacterial spores. Biochemical Journal 54:205–209
    [Google Scholar]
  16. Strange R., Dark F. 1957; A cell-wall lytic enzyme associated with spores of Bacillus species. Journal of General Microbiology 16:236–249
    [Google Scholar]
  17. Vary J. C. 1973; Germination of Bacillus megaterium spores after various extraction procedures. Journal of Bacteriology 116:797–802
    [Google Scholar]
  18. Warth A. 1972; Action of spore lytic enzymes on the cortex. In Spores V pp. 28–34 Halvorson H. O., Hanson R., Campbell L. L. Edited by Washington, D.C.: American Society for Microbiology.;
    [Google Scholar]
  19. Wyatt L., Waites W. M. 1974; The effect of sodium hydroxide or dithiothreitol-urea on spores of germination mutants of Clostridium bifermentans. Journal of General Microbiology 84:391–394
    [Google Scholar]
/content/journal/micro/10.1099/00221287-126-1-37
Loading
/content/journal/micro/10.1099/00221287-126-1-37
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