Dipeptidyl peptidase IV of Streptococcus anginosus: purification and characterization.

We found that N-unblocked nine p-nitroanilde derivatives of amino acids or peptides were hydrolyzed by the crude cell extracts of Streptococcus anginosus NCTC 10713. Then dipeptidyl peptidase IV was purified 323-fold by the procedures including ammonium sulfate concentration, anion exchange chromatography (twice), gel filtration (twice), hydrophobic interaction chromatography, and isoelectric focusing. The molecular weight was calculated as 84 kDa, and the isoelectric point was 4.9. The enzyme hydrolyzed mainly dipeptides containing proline residues at P1 position. It was strongly inhibited by serine enzyme inhibitors. General protease inhibitors, metal chelators, thiol alkylating agent, reducing agent, and several metal ions had no effect on the enzyme activity. Optimum pH for the activity was found at 7.0. The enzyme was mostly inactivated by heating at 50 degrees C for 15 min.