1887

Abstract

Summary

Ten isolates of expressing different degrees of toxigenicity and virulence in an animal model were assayed for the production of proteolytic enzymes by various methods. All strains demonstrated some acitivity in one or more of the assay systems. There was no direct correlation between toxigenic status and enzyme production. However, those strains known to be highly virulent in a hamster model were the most proteolytic. The most commonly detected enzyme was cell associated, and its substrate specificity suggested it was a trypsin-like enzyme. Initial purification of the enzyme from strain VPI 10463 gave a 10% yield with a 14-fold increase in purity. Inhibition studies on this preparation indicated that the enzyme was a thiol protease. The enzyme has pH and temperature optima of 7–5 and 37°C, respectively. These characteristics suggest that the enzyme is more related to clostripain, the thiol clostridio-peptidase of , than to trypsin. Whilst the role of this enzyme remains unclear, it is possible that it may be a contributory factor in the virulence of the organism as described for other clostridial infections.

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1992-05-01
2022-05-20
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References

  1. George R. H., Symonds J. M., Dimock F. Identification of Clostridium difficile as a cause of pseudomembranous colitis. Br Med J 1978; 1:695
    [Google Scholar]
  2. Larson H. E., Price A. B., Honour P., Bordello S. P. Clostridium difficile and the aetiology of pseudomembranous colitis. Lancet 1978; 1:1063–1066
    [Google Scholar]
  3. Bordello S. P., Larson H. E. Antibiotics and pseudomembranous colitis. J Antimicrob Chemother 1981; 7: Suppl A 53–62
    [Google Scholar]
  4. Taylor N. S., Thome G. M., Bartlett J. G. Comparison of two toxins produced by Clostridium difficile. Infect Immun 1981; 34:1036–1043
    [Google Scholar]
  5. Lyerly D. M., Krivan H. C., Wilkins T. D. Clostridium difficile: its disease and toxins. Clin Microbiol Rev 1988; 1:1–18
    [Google Scholar]
  6. Bordello S. P., Welch A. R., Barclay F. E., Davies H. A. Mucosal association by Clostridium difficile in the hamster gastrointestinal tract. J Med Microbiol 1988; 25:191–196
    [Google Scholar]
  7. Bordello S. P., Davies H. A., Barclay F. E. Detection of fimbriae amongst strains of Clostridium difficile. FEMS Microbiol Lett 1988; 49:65–67
    [Google Scholar]
  8. Bordello S. P., Davies H. A., Kamiya S., Reed P. J., Seddon S. Virulence factors of Clostridium difficile. Rev Infect Dis 1990; 12: Suppl 2S185–S191
    [Google Scholar]
  9. Seddon S. V., Hemingway I., Bordello S. P. Hydrolytic enzyme production by Clostridium difficile and its relationship to toxin production and virulence in the hamster model. J Med Microbiol 1990; 31:169–174
    [Google Scholar]
  10. Steffen E. K., Hentges D. J. Hydrolytic enzymes of anaerobic bacteria isolated from human infections. J Clin Microbiol 1981; 14:153–156
    [Google Scholar]
  11. Maclennon J. D. The histotoxic clostridial infections of man. Bacteriol Rev 1962; 26:177–276
    [Google Scholar]
  12. Seddon S. V., Bordello S. P. A chemically defined and minimal medium for Clostridium difficile. Lett Appl Microbiol 1989; 9:237–239
    [Google Scholar]
  13. Bradford M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72:248–254
    [Google Scholar]
  14. Van Steenbergen T. J. M., de Graaff J. Proteolytic activity of black pigmented bacteroides strains. FEMS Microbiol Lett 1986; 33:219–222
    [Google Scholar]
  15. Gibson S. A. W., Macfarlane G. T. Studies on the proteolytic activity of Bacteroides fragilis. J Gen Microbiol 1988; 134:19–27
    [Google Scholar]
  16. Kamiya S., Reed P. J., Borriello S. P. Analysis of purity of Clostridium difficile toxin A derived by affinity chromatography on immobilized bovine thyroglobulin. FEMS Microbiol Lett 1988; 56:331–336
    [Google Scholar]
  17. Borriello S. P., Ketley J. M., Mitchell T. J. Clostridium difficile– a spectrum of virulence and analysis of putative virulence determinants in the hamster model of antibiotic-associated colitis. J Med Microbiol 1987; 24:53–64
    [Google Scholar]
  18. Corthier G., Muller M. C., Elmer G. W., Lucas F., Dubos-Ramare F. Relationship between proteolytic activities, and production of toxin A during Clostridium difficile induced cecitis in gnotobiotic mice. In Dougherty S. H. (eds) Microecology and therapy vol 19 Herbom-Dill: Institute for Microecology; 1990217
    [Google Scholar]
  19. Kamiya S., Borriello S. P. A non-haemagglutinating form of Clostridium difficile toxin A. J Med Microbiol 1992; 36:190–197
    [Google Scholar]
  20. Lyerly D. M., Carrig P. E., Wilkins T. D. Susceptibility of Clostridium difficile toxins A and B to trypsin and chymotrypsin. Microb Ecol Health Dis 1989; 2:219–221
    [Google Scholar]
  21. Patterson-Curtis S. I., Johnson E. A. Regulation of neurotoxin and protease formation in Clostridium botulinum Okra B and Hall A by arginine. Appl Environm Microbiol 1989; 55:1544–1548
    [Google Scholar]
  22. Mitchell W. M., Harrington W. F. Clostripain. In Boyer P. D. (ed) The enzymes vol 3 New York: Academic Press; 1971699–719
    [Google Scholar]
  23. Mitchell W. M., Harrington W. F. Purification and properties of Clostripeptidase B (Clostripain). J Biol Chem 1968; 243:4683–4692
    [Google Scholar]
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