Characterization of a trypsin-like protease from the bacterium Bacteroides gingivalis isolated from human dental plaque.

A trypsin-like, membrane-bound protease from Bacteroides gingivalis was solubilized by Triton X-100 and partially purified by a combination of DEAE-Sepharose and aminophenylmercuric Sepharose chromatography, by taking advantage of the thiol group on the enzyme. The purified enzyme hydrolysed the synthetic substrates benzoyl-L-arginine-p-nitroanilide (L-BAPA), benzoyl-D,L-arginine-beta-naphthylamide (BANA) and tosyl-L-arginine methyl ester, as well as bovine serum albumin and ovalbumin, but not tosyl-L-lysine methyl ester. The enzyme activity was enhanced by SH-reagents and was inhibited to different degrees by SH-inhibitors, chelators and microbial low-molecular-weight inhibitors such as leupeptin, antipain and chymostatin. These microbial inhibitors could be of practical use as ligands for affinity chromatography for further purification. The possible involvement of the protease in periodontal diseases is also discussed.