pH and Temperature Optimization for Enzymatic Activity

Optimum pH requirement is very important for an enzyme-substrate reaction, so broader pH range 5.0–12.0 was investigated. The pH values were obtained with the addition to different buffer solutions: sodium acetate buffer, pH 5.0; phosphate buffer, pH 6.0–8.0; and glycine–NaOH buffer, pH 9.0–12.0. The reaction mixture was incubated at optimum temperature and activity was determined by DNS method. pH stability was determined by incubating enzyme in the above-mentioned buffers at 65°C for 1 h and, in terms of temperature and temperature stability after incubating enzyme at a different temperature range (30–95°C) for 1 h, residual amylase activity was measured under assay conditions.