CAPNOCYTOPHAGA GINGIVALIS AMINOPEPTIDASE - A POTENTIAL VIRULENCE FACTOR

The production and properties of an aminopeptidase from Capnocytophaga gingivalis were studied. C. gingivalis was grown in continuous cultur e 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 (0.6 mu(rel)) TLPase specific activity was low an d found exclusively as cell-bound activity; at low growth rates (0.037 5 mu(rel)), specific activity was high and 26% was found as extracellu lar activity. In contrast, aminopeptidase specific activity was highes t at 0.3 mu(rel) and the ratio of cell-bound to extracellular activity was relatively constant at all growth rates. Only about 5% of the tot al activity was extracellular. The aminopeptidase, which has a wide sp ecificity towards artificial substrates, was purified to homogeneity, as judged by SDS-PAGE, from the supernatant fluid of cells grown in co ntinuous 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 an d showed a requirement for magnesium or calcium ions. The N-terminal s equence 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 nutritio n and pathogenesis of C. gingivalis in the human oral cavity.