Open AccessIsolation and properties of a tripeptidyl peptidase from a periodontal pathogen Prevotella nigrescens
Author(s) -
Fujimura Setsuo,
Ueda Ohmi,
Shibata Yukinaga,
Hirai Kaname
Publication year - 2003
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/s0378-1097(03)00048-x
Subject(s) - isoelectric point , isoelectric focusing , enzyme , biochemistry , chromatography , prevotella , size exclusion chromatography , chemistry , serine , molecular mass , amino acid , residue (chemistry) , bacteria , biology , genetics
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°C for 5 min and inhibited significantly by specific serine enzyme inhibitors.