Human pancreatic alpha-amylase. II. Effects of pH, substrate and ions on the activity of the enzyme.
Purified human pancreatic alpha-amylase (alpha-1,4-glucan 4-glucano-hydrolase, EC 3.2.1.1) was found to be stable over a wide range of pH values (5.0 to 10.5) with an optimal pH for the enzymatic activity of 7.0. The Michaelis constant of the enzyme at optimal pH and assay conditions was found to be 2.51 mg per ml for soluble starch. Halide ions were required for the activity of the enzyme whereas sulfate and nitrate were not. The order of effectiveness of activation was found to be: Cl- greater than Br- greater than I- greater than F-. Calcium and magnesium were activators at concentrations of 0.001M and 0.005M, respectively, but exhibited inhibitory effects at concentrations higher than 0.005M. At 0.01M ethylenediamine tetraacetic acid (EDTA) concentration the enzymatic activity upon seven min incubation, was inhibited up to 96%. The inhibition of EDTA and calcium could be reversed upon addition of calcium and EDTA, respectively.
Human pancreatic alpha-amylase. II. Effects of pH, substrate and ions on the activity of the enzyme. Purified human pancreatic alpha-amylase (alpha-1,4-glucan 4-glucano-hydrolase, EC 3.2.1.1) was found to be stable over a wide range of pH values (5.0 to 10.5) with an optimal pH for the enzymatic activity of 7.0. The Michaelis constant of the enzyme at optimal pH and assay conditions was found to be 2.51 mg per ml for soluble starch. Halide ions were required for the activity of the enzyme whereas sulfate and nitrate were not. The order of effectiveness of activation was found to be: Cl- greater than Br- greater than I- greater than F-. Calcium and magnesium were activators at concentrations of 0.001M and 0.005M, respectively, but exhibited inhibitory effects at concentrations higher than 0.005M. At 0.01M ethylenediamine tetraacetic acid (EDTA) concentration the enzymatic activity upon seven min incubation, was inhibited up to 96%. The inhibition of EDTA and calcium could be reversed upon addition of calcium and EDTA, respectively.