N-α-benzoyl-DL-arginine ρ-nitroanilide hydrolase (BAρNAase) was partially purified and characterized from the cotyledons of Canavalia lineata. The overall purification and final recovery were 77.5-fold and 7%, respectively. The apparent mol wt of t...
N-α-benzoyl-DL-arginine ρ-nitroanilide hydrolase (BAρNAase) was partially purified and characterized from the cotyledons of Canavalia lineata. The overall purification and final recovery were 77.5-fold and 7%, respectively. The apparent mol wt of the native enzyme was estimated as 200 kD from the gel filtration and this enzyme was revealed as one of the gelatin-hydrolyzed proteases by activity staining with gelatin-containing gel. The optimum pH for the hydrolysis of N-α-benzoyl-DL-arginine ρ-nitroanilide (BAρNAase) was about 9.5. This enzyme catalyzed the hydolysis of BAρNA with Vmax of 15.5 unit/mg and Km value of 1.6 mM which were different from them of bovine-pancreatic trypsin. The catalytic activity of this enzyme was strongly inhibited by phenylmethanesulfonyl fluoride (PMSF) but insensitive to aprotinin, indicating that it was a serine proteinase but not a trypsin-like one. And this enzyme was slightly stimulated by Ca^2+ and mg^2+ but remarkably inhibited by Mn^2+, HG^2+ and Zn^2+.