Substrate-Selective Activation of Histidine-Modified Porcine Pancreatic α-Amylase by Chloride Ion

Abstract

Porcine pancreatic α-amylase (1, 4-α-D-glucan glucanohydrolase) [EC 3.2.1.1] has both amylase activity (hydrolysis of α-1, 4-D-glucoside bond of starch) and maltosidase activity (hydrolysis of p-nitrophenyl-α-D-maltoside to p-nitrophenol and maltose). By the modification of histidine residues of porcine pancreatic α-amylase with diethylpyrocarbonate (DEP), both amylase and maltosidase activities were decreased in the absence of chloride ion. In the presence of chloride ion, however, maltosidase activity of the modified enzyme was increased to more than 260% of that of the native enzyme, whereas amylase activity was decreased to less than 15% of the native enzyme. Since the chloride ion binding site is part of the active site loop [Buisson et al. (1987) Food Hydrocolloids 1, 399-406 and Buisson et al. (1987) EMBO J. 6, 3909-3916], the special arrangements of both catalytic and modified histidine residues induced by the chloride ion binding would enhance only the maltosidase activity of the histidine-modified enzyme.