Isolation and properties of a tripeptidyl peptidase from a periodontal pathogen Prevotella nigrescens

Prolyltripeptidyl amino peptidase activity was found in a crude extract of Prevotella nigrescens and this Enzyme was purified by procedures including concentration with ammonium sulfate, ion exchange chromatography, gel filtration, and isoelectric focusing. This peptidase hydrolyzed Ala-Ala-Pro-p-nitroanilide as well as Ala-Phe-Pro-p-nitroanilide. Furthermore, several p-nitroanilide derivatives of dipeptides with a proline residue in the second position from the amino-terminal end (Xaa-Pro) were also cleaved detectably. The molecular mass of this tripeptidase was calculated as 56 kDa and its isoelectric point was 5.8. The Enzyme was inactivated completely by heating at 60 degrees C for 5 min and inhibited significantly by specific serine Enzyme inhibitors.