1887

Abstract

a bacterial species frequently associated with disease periodontal sites, is known to possess trypsin-like activity. The present study was undertaken to determine the major characteristics of this activity. The trypsin-like activity was mainly found on the surface of the bacteria and could be solubilized with a zwitterionic detergent (Zwittergent 3-14). Using -a-benzoyl-DL-arginine--nitroanilide as substrate, the optimum pH was between 7·5 and 8·5 and the optimum temperature was 35°C. The evidence suggests that the enzyme is a serine protease since it was strongly inhibited by diisopropylfluorophosphate (DFP), -a--tosyl-L-lysine chloromethyl ketone hydrochloride, leupeptin and antipain. The trypsin-like enzyme cleaved numerous chromogenic synthetic peptides containing either an arginine or lysine bond, but could not hydrolyse native proteins including casein, gelatin and BSA. Incubation of a cell envelope extract of the presence of [H]DFP, which is known to bind irreversibly to serine proteases, labelled two bands at 70 and 81 kDa following SDS-PAGE (under reducing conditions) and fluorography. It is suggested that the trypsin-like enzyme may be mainly involved in the degradation of small peptides resulting from hydrolysis of larger proteins by other oral bacteria.

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1995-04-01
2024-04-24
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References

  1. Aphale J. S., Strohl W. R. 1993; Purification and properties of an extracellular aminopeptidase from Streptomjces lividans1326. J Gen Microbiol 139:417–424
    [Google Scholar]
  2. Dzink J. L., Smith C., Socransky S. S. 1988; The predominant cultivable microbiota of active and inactive lesions of destructive periodontal diseases. J Clin Periodontol 15:316–323
    [Google Scholar]
  3. Grenier D., Mayrand D. 1993 Proteinases. In Biology of the species Porphyromonasgingivalis pp 227–243 Edited by Shah H. N., Mayrand D., Genco R. J. Boca Raton: CRC Press.;
    [Google Scholar]
  4. Holt S. C., Bramanti T. E. 1991; Factors in virulence expression and their role in periodontal disease pathogenesis. Crit Ren Oral Biol Med 2:177–281
    [Google Scholar]
  5. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  6. Lai C.-H., Listgarten M. A., Shirakawa M., Slots J. 1987; Bacteroides forsythus in adult gingivitis and periodontitis. Oral Microbiol Immunol 2:152–157
    [Google Scholar]
  7. Laughon B. E., Syed S. A., Loesche W. J. 1982; API-ZYM system for identification of Bacteroides sp., Capnocytophaga sp., and spirochetes of oral origin. J Clin Microbiol 15:97–102
    [Google Scholar]
  8. Loesche W. J., Bretz W. A., Kerschensteiner D., Stoll J., Socransky S. S., Hujoel P., Lopatin D. E. 1990; Development of a diagnostic test for anaerobic periodontal infections based on plaque hydrolysis of benzoyl-dl-arginine-naphthylamide. J Clin Microbiol 28:1551–1559
    [Google Scholar]
  9. Mayrand D., McBride B. C. 1980; Ecological relationships of bacteria involved in a simple, mixed anaerobic infection. Infect Immun 27:4450
    [Google Scholar]
  10. Ohta K., Makinen K. K., Loesche W. J. 1986; Purification and characterization of an enzyme from Treponema denticola capable of hydrolyzing synthetic trypsin substrates. Infect Immun 53:213–220
    [Google Scholar]
  11. Shah H. N., Williams R. A. D. 1982; Dehydrogenase patterns in the taxonomy of Bacteroides . J Gen Microbiol 128:2955–2965
    [Google Scholar]
  12. Slots J. 1981; Enzymatic characterization of some oral and non oral Gram negative bacteria with the API ZYM system. J Clin Microbiol 14:288–294
    [Google Scholar]
  13. Socransky S. S., Haffajee A. D. 1991; Microbial mechanisms in the pathogenesis of destructive periodontal diseases: a critical assessment. J Periodontal Res 26:195–212
    [Google Scholar]
  14. Socransky S. S., Haffajee A. D. 1992; The bacterial etiology of destructive periodontal disease: current concepts. J Periodontol 63:322–331
    [Google Scholar]
  15. Tanner A. C. R., Strzempko M. N., Belsky C. A., McKinley G. A. 1985; API ZYM and API An-Ident reactions of fastidious oral Gram-negative species. J Clin Microbiol 22:333–335
    [Google Scholar]
  16. Tanner A. C. R., Listgarten M. A., Ebersole J. L., Strzempko M. N. 1986; Bacteroides forsythus sp. nov., a slow-growing, fusiform Bacteroides sp. from the oral cavity. Int J Syst Bacteriol 36:213–221
    [Google Scholar]
  17. Williams R. C. 1990; Periodontal disease. New Engl J Med 322:373–382
    [Google Scholar]
  18. Wyss C. 1989; Dependence of proliferation of Bacteroides forsythus on exogenous N-acetylmuramic acid. Infect Immun 57:1757–1759
    [Google Scholar]
  19. Yamashita Y., Toyoshima K., Yamazaki M., Hanada N., Takehara T. 1990; Purification and characterization of alkaline phosphatase of Bacteroides gingivalis 381. Infect Immun 58:2882–2887
    [Google Scholar]
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