1887

Abstract

strain W50 was grown in batch and continuous culture on complex medium with haemin. In batch culture, cell-bound levels of trypsin-like protease (EC 3.4.21.4), alkaline phosphatase (EC 3.1.3.1) and -acetyl--glucosaminidase (EC 3.2.1.30) increased during the exponential phase of growth. These enzyme activities were also detected in extracellular vesicles and in extracellular soluble forms in the supernatant fluid, but in lower amounts per unit biomass compared to cell-bound levels. In continuous culture, at high relative growth rates (0·7–0·9 ), the highest proportions of enzyme activities were cell-bound. In contrast, at low relative growth rates (0·1–0·2 ), highest enzyme levels were detected in the extracellular vesicle fraction. Levels of extracellular soluble enzymes were always low compared to cell-bound or extracellular vesicle levels, but were highest at low relative growth rates. All three enzymes appeared to be relatively stable in their soluble forms. Vesicle production appeared to be associated with actively growing cells but was influenced by growth rate. The results are consistent with the hypothesis that cell-bound periplasmic enzymes are encapsulated into vesicles which are subsequently released by the cells. Therefore, levels of total extracellular enzyme (extracellular vesicle plus extracellular soluble) may depend on the rate of vesicle formation superimposed on the rates of production of periplasmic enzymes in the cell.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-135-3-557
1989-03-01
2024-05-08
Loading full text...

Full text loading...

/deliver/fulltext/micro/135/3/mic-135-3-557.html?itemId=/content/journal/micro/10.1099/00221287-135-3-557&mimeType=html&fmt=ahah

References

  1. Cheng K.-J., Ingram J.M., Costerston J.W. 1970; Alkaline phosphatase localization and sphaeroplast formation ofPseudomonas aeruginosa . Canadian Journal of Microbiology 16:1319–1324
    [Google Scholar]
  2. Coykendall A.L., Kaczmarek F.S., Slots J. 1980; Genetic heterogeneity inBacteroides asac-charolyticus (Holdeman and Moore, 1970) Finegold and Barnes, 1977 (Approved Lists, 1980) and proposal of Bacteroides gingivalis sp. nov. and Bacteroides macacae (Slots and Genco) comb. nov. International Journal of Systematic Bacteriology 30:559–564
    [Google Scholar]
  3. Eaves G., Greenman J., Holland K.T. 1979; Decay rates of exocellular enzymes produced by bacteria - correction factor of production rates in chemostat cultures. FEMS Microbiology Letters 6:333–336
    [Google Scholar]
  4. Fujimura S., Nakamura T. 1987; Isolation and characterization of a protease fromBacteroides gingivalis . Infection and Immunity 55:716–720
    [Google Scholar]
  5. Greenman J., Holland K.T., Cunliffe W.J. 1981; Effects of glucose concentration on biomass, maximum specific growth rate and extracellular enzyme production by three species of cutaneous propionibacteria grown in continuous culture. Journal of General Microbiology 127:371–376
    [Google Scholar]
  6. Grenier D., Mayrand D. 1987; Functional characterization of extracellular vesicles produced byBacteroides gingivalis . Infection and Immunity 55:111–117
    [Google Scholar]
  7. Holland K.T., Greenman J., Cunliffe W.J. 1979; Growth of cutaneous propionibacteria on synthetic medium; growth yields and exoenzyme production. Journal of Applied Bacteriology 47:383–394
    [Google Scholar]
  8. Laughon B.E., Syed S.A., Loesche W.J. 1982; API ZYM system for identification ofBacteroidesspp., Capnocytophaga spp., and spirochetes of oral origin. Journal of Clinical Microbiology 15:97–102
    [Google Scholar]
  9. McKee A.S., McDermid A.S., Baskerville A., Dowsett A.B., Ellwood D.C., Marsh P.D. 1986; Effect of hemin on the physiology and virulence ofBacteroides gingivalis . Infection and Immunity 52:349–355
    [Google Scholar]
  10. Neu H.C., Heppel L.A. 1965; The release of enzymes fromEscherichia coli by osmotic shock and during the formation of spheroplasts. Journal of Biological Chemistry 240:3685–3692
    [Google Scholar]
  11. Shah H.N., Williams R.A.D., Bowden G.D., Hardie J.M. 1976; Comparison of the biochemical properties ofBacteroides melaninogenicus from human dental plaque and other sites. Journal of Applied Bacteriology 41:473–492
    [Google Scholar]
  12. Silhavy T.J., Benson S.A., Emr S.D. 1983; Mechanism of protein localization. Microbiological Reviews 41:313–344
    [Google Scholar]
  13. Slots J. 1982; Importance of black-pigmentedBacteroides in human periodontal disease. In Host- Parasite Interactions in Periodontal Diseases pp Genco R. J., Mergenhagen S. E. Edited by Washington, DC: American Society for Microbiology;
    [Google Scholar]
  14. Slots J., Dahlen G. 1985; subgingival microorganisms and virulence factors in periodontitis. Scandinavian Journal of Dental Research 93:119–127
    [Google Scholar]
  15. Slots J., Genco R.J. 1984; Black-pigmentedBacteroides species, Capnocytophaga species, and Actinobacillus actinomycetemcomitans in human periodontal disease: virulence factors in colonization, survival and tissue destruction. Journal of Dental Research 63:412–421
    [Google Scholar]
  16. Slots J., Gibbons R.J. 1978; Attachment ofBacteroides melaninogenicus subspecies asaccharolyti-cus to oral surfaces and its possible role in colonization of the mouth and of periodontal pockets. Infection and Immunity 19:254–264
    [Google Scholar]
  17. Smalley J.W., Birss A.J. 1987; Trypsin-like enzyme activity of the extracellular membrane vesicles ofBacteroides gingivalis W50. Journal of General Microbiology 133:2883–2894
    [Google Scholar]
  18. Tempest D.W. 1970; The continuous cultivation of microorganisms. 1. Theory of the chemostat. Methods in Microbiology 2:259–276
    [Google Scholar]
  19. Tempest D.W. 1976; The concept of‘relative growth rate’: its theoretical basis and practical application. In Continuous Culture vol.6, Applications and New Fields pp 349–352 Dean A. C. R., Ellwood D. C., Evans C. G. T., Melling J. Edited by Chichester: Ellis Horwood;
    [Google Scholar]
  20. VanWinkelhoff A.J., VanSteenbergen T.J.M., Kippuw N., deGraaff J. 1985; Further characterization ofBacteroides endodontalis as asac- charolytic black-pigmented Bacteroides species from the oral cavity. Journal of Clinical Microbiology 22:75–79
    [Google Scholar]
  21. Woo D.D.L., Holt S.C., Leadbetter E.R. 1979; Ultrastructure ofBacteroides species: Bacteroides asaccharolyticus, Bacteroides fragilis, Bacteroides melaninogenicus subspecies melaninogenicus and Bacteroides melaninogenicus subspecies intermedius . Journal of Infectious Diseases 139:534–546
    [Google Scholar]
  22. Yoshimura F., Nishikata M., Suzuki T., Hoover C.I., Newbrun E. 1984; Characterization of a trypsin-like protease from the bacteriumBacteroides gingivalis isolated from human dental plaque. Archives of Oral Biology 29:559–564
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-135-3-557
Loading
/content/journal/micro/10.1099/00221287-135-3-557
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