1887

Abstract

The production and properties of an aminopeptidase from were studied. was grown in continuous culture over a range of dilution rates and the cell-bound and extracellular levels of aminopeptidase and trypsin-like protease (TLPase) measured. At high growth rates (0.6μ) TLPase specific activity was low and found exclusively as cell-bound activity; at low growth rates (0.0375 μ), specific activity was high and 26% was found as extracellular activity. In contrast, aminopeptidase specific activity was highest at 0.3 μ and the ratio of cell-bound to extracellular activity was relatively constant at all growth rates. Only about 5% of the total activity was extracellular. The aminopeptidase, which has a wide specificity towards artificial substrates, was purified to homogeneity, as judged by SDS-PAGE, from the supernatant fluid of cells grown in continuous culture in a tryptone/glucose/thiamine medium. The enzyme has a molecular mass of 61 kDa, a pI of 6.3, a pH optimum close to 7.5 and showed a requirement for magnesium or calcium ions. The N-terminal sequence of the first 10 amino acids (Asp-Val-Asn-Met-Leu-Trp-Tyr-Val-x-Arg…) showed no similarity to any published sequence. This enzyme in its cell-bound or extracellular form may be important in the nutrition and pathogenesis of in the human oral cavity.

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1995-12-01
2021-05-14
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References

  1. Andersson C, Linder L.E., Sund M., Lönnies H. 1992; Purification and characterization of an aminopeptidase from Streptococcus mitis ATCC 903.. Curr Microbiol 25:261–267
    [Google Scholar]
  2. Ando K. 1980; Collagenase, dipeptidylpeptidase IV and cathepsin D activities in gingival fluid and whole saliva from patients with periodontal disease.. Jap J Periodont 22:387–402
    [Google Scholar]
  3. Ben-Basset A., Bauer K., Sheng-Yung C., Myambo C., Boosman A., Chang S. 1987; Processing of the initiation methionine from proteins; properties of the Escherichia coli methionine aminopeptidase and its gene structure.. J Bacteriol 169:751–757
    [Google Scholar]
  4. Bickel M., Cimasoni G. 1986; Recent advances in gingival crevicular fluid research.. In The Borderland Between Caries and Periodontal Disease III. Lehner T., Cimisoni G. Edited by Geneva: Médecine et Hygiene;
    [Google Scholar]
  5. Bradford M.M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.. Anal Biochem 72:248–254
    [Google Scholar]
  6. Cueva R., Garcia-Alvarez N., Suarez-Rendueles P. 1989; Yeast vacuolar aminopeptidase yscI. Isolation and regulation of the APE1 (LAP4) structural gene.. FEBS Lett 259:125–129
    [Google Scholar]
  7. D’Amato R.F., Eriquez L.A., Tomfohrde K.M., Singerman E. 1978; Rapid identification of Neisseria gonorrhoeae and Neisseria meningitidis by using enzymatic profiles.. J Clin Microbiol 7:77–81
    [Google Scholar]
  8. Fricke B., Aurich H. 1993; Periplasmic aminopeptidases in Acinetobacter calcoaceticus and Pseudomonas aeruginosa. . J Basic Microbiol 33:291–299
    [Google Scholar]
  9. Green S.S., Goldberg H.S., Blenden D.C. 1967; Enzyme patterns in the study of Leptospira. . Appl Microbiol 15:1104–1113
    [Google Scholar]
  10. 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.. J Gen Microbiol 127:371–376
    [Google Scholar]
  11. Holdeman LV., Moore W.E., Cato E.P., Burmeister J.A., Palcanis K.G., Ranney R.R. 1985; Distribution of Capnocjtophaga in periodontal microfloras.. J Periodont Res 20:475–483
    [Google Scholar]
  12. Holland K.T., Greenman J., Cunliffe W.J. 1979; Growth of Propionibacteria on synthetic medium: growth yields and exoenzyme production.. J Appl Bact 47:383–394
    [Google Scholar]
  13. Hopsu-Havu V.K., Mäkinen K.K., Glenner G.G. 1966; Purification of a mammalian peptidase selective for N-terminal arginine and lysine residues: aminopeptidase B.. Arch Biochem Biophjs 114:557–566
    [Google Scholar]
  14. Ingham E., Holland K.T., Gowland G., Cunliffe W.J. 1983; Studies of the extracellular proteolytic activity produced by Propionibacterium acnes. . J Appl Bact 54:263–271
    [Google Scholar]
  15. Irving J.T., Socransky S.S., Tanner A.C.R. 1978; Histological changes in experimental periodontal disease in rats mono-infected with Gram-negative organisms.. J Periodont 13:326–332
    [Google Scholar]
  16. Kumano N., Sugawara S. 1992; Ubenimex (Bestatin), an aminopeptidase inhibitor, modulates protein kinase C in K562 cells.. J Biol Regulat Homeostat Agents 6:116–120
    [Google Scholar]
  17. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 227:680–685
    [Google Scholar]
  18. Leadbetter E.R., Holt S.C., Socransky S.S. 1979; Capnocytophaga: new genus of Gram-negative gliding bacteria. I. General characteristics, taxonomic considerations and significance.. Arch Microbiol 122:9–16
    [Google Scholar]
  19. Mäkinen K.K., Syed S.A., Loesche W.J., Mdkinen P.-L. 1988; Proteolytic profile of Treponema vincentii ATCC 35580 with special reference to collagenolytic and arginine aminopeptidase activity.. Oral Microbiol Immunol 3:121–128
    [Google Scholar]
  20. Marsh P.D., Hunter J.R., Bowden G.H., Hamilton I.R., McKee A.S., Hardie J.M., Ellwood D.C. 1983; The influence of growth rate and nutrient limitation on the microbial composition and biochemical properties of a mixed culture of oral bacteria grown in a chemostat.. J Gen Microbiol 129:755–770
    [Google Scholar]
  21. Mashimo P.A., Yamamoto Y., Slots J., Park 8.H., Genco R.J. 1983; The periodontal microflora of juvenile diabetics: culture, immunofluorescence and serum antibody studies.. J Periodontol 54:420–430
    [Google Scholar]
  22. Minhas T., Greenman J. 1989; Production of cell-bound and vesicle-associated trypsin-like protease, alkaline phosphatase and N-acetyl-fS-glucosaminidase by Bacteroides gingivalis strain W50.. J Gen Microbiol 135:557–564
    [Google Scholar]
  23. Nakamura M., Slots J. 1982; Aminopeptidase activity of Capnocjtophaga. . J Periodont Res 17:597–603
    [Google Scholar]
  24. Newman M.G., Socransky S.S., Savitt E.D., Crawford A. 1976; Studies in the microbiology of periodontosis.. J Periodontol 47:373–379
    [Google Scholar]
  25. Shibata Y., Fujimura S., Nakamura T. 1992; Isolation and characterization of enzymes hydrolyzing chymotrypsin synthetic substrate (enzyme 1) and trypsin synthetic substrate (enzyme 2) from the envelope fraction of Capnocytophagagingivalis. . Med Microbiol Immunol 181:107–115
    [Google Scholar]
  26. Slots J. 1976; The predominant cultivable organisms in juvenile periodontitis.. Scand J Dent Res 84:1–10
    [Google Scholar]
  27. Slots J. 1982; Importance of black-pigmented Bacteroides in human periodontal disease.. Etiol Microb Spec 2:27–45
    [Google Scholar]
  28. Spratt D.A. 1994 A study of aminopeptidase and other hydrolytic ensymesproduced by Capnocytophaga gingivalis. PhD Thesis University of the West of England,Bristol,UK.:
    [Google Scholar]
  29. Tempest D.W. 1970; The continuous cultivation of microorganisms. I. Theory of the chemostat.. Methods Microbiol 2:259–276
    [Google Scholar]
  30. Umezawa H. 1982; Low molecular weight enzyme inhibitors of microbial origin.. Annu Rev Microbiol 36:75–99
    [Google Scholar]
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