The distribution and properties of some hydrolytic enzymes from Porphyromonas gingivalis W50

The production, distribution (as cell-bound, extracellular or associated with vesicles) and properties of four hydrolytic enzymes from Porphyromonas gingivalis were studied. During batch culture, enzymes appeared as cell bound until 72 h when extracellular trypsin-like protease and glycylprolyl dipeptidase were found. Alkaline phosphatase and N-acetyl-beta-glucosaminidase continued to be mainly cell-bound until 96 and 144 h post-inoculation, respectively. The pH optima for trypsin-like protease and dipeptidase activities were 7.5 and 8.0, respectively. Activity was most stable between pH 6.5 and 9.0 for the trypsin-like protease and pH 8.0 for dipeptidase. For phosphatase and N-acetyl-beta-glucosaminidase, double peaks of activity occurred at pH 8.0, 10.5 and 6.5, 7.5, respectively. Phosphatase was most stable at pH 7.0, whilst N-acetyl-beta-glucosaminidase was stable over a wide range of pH from 5.0-10.0. Molecular weight estimation by gel filtration gave 170 and 65 kD for trypsin-like protease, 200 and 24 kD for glycylprolyl dipeptidase, 125 and 24 kD for phosphatase, and 22 kD for N-acetyl-beta-glucosaminidase.