Abstract: The purpose of this study was to optimize α-amylase inhibitors by response surface methodology with industrial α-amylase instead of pancreatic α-amylase and to investigate α-amylase reactive system included pH value, incubation temperature, and proportion in soluble amylum and α-amylase by single factor analysis and response surface analysis. The α-amylase activity was examed by 3, 5-dinitrosalicylic acid colorimetric assays (Bemfeld method) . The final conditions of reactive system were pH6.6 value, 55℃ and proportion of 1∶1.5. Results showed that the optimize method of α-amylase inhibitors was stable, reliable and well repeated. It could be used to screen the inhibitors of α-amylase.