Purification and Characterization of Rice Embryo BAPAase (Benzoyl-L-arginine p-nitroanilide Hydrolase)

Publisher Website
Google Scholar
Abstract
Benzoyl-L-arginine p-nitroanilide hydrolase (BAPAase), which has both endopeptidase and carboxypeptidase activity toward the Arg-X or Lys-X bond of small peptides [Shibata and Doi (1984) Plant & Cell Physiol. 25: 1421], was purified from rice embryos by ammonium sulfate and polymin fractionations and by ion exchange, gel exclusion and hydrophobic chromatographies. The purified enzyme was homogeneous when analyzed by polyacrylamide gel electrophoresis. It was unstable in the absence of surface-active reagents such as Triton X-100. Maximum activity for benzoyl-Larginine p-nitroanilide (L-BAPA) or carboxypeptidase activity toward butoxycarbonyl-Gly-Lys-Leu was obtained at pH 9.0. L-BAPA at high concentrations inhibited the enzyme's activity. Di-isopropyl phosphofluoridate, N-tosyl-L-lysine chloromethyl ketone, leupeptin and antipain, which are specific inhibitors of trypsin, inhibited BAPAase activity, but soybean and rice bran trypsin inhibitor had no effect on it. Sulfhydryl reagents strongly inhibited the BAPAase activity.

This publication has 1 reference indexed in Scilit: