Plant alpha-amylase inhibitors show great potential as tools to engineer resistance of crop plants against pests. They are also drug-design targets for treatment of diabetes and digestion disorderes.These inhibitors also known as sensitizing agents in human. The numerous form of alpha -amylase inhibitors was reported. In this study alpha-amylase inhibitor was extracted from Iranian wheat cultivar (Triticum aestivum v zarrin), precipitated and purified by anion exchange fast protein liquid chromatography. Electrophoresis of purified protein showed 0.66 relative mobility .Total hydrolytic activity of human salivary and bacillus subtilis alpha -amylase were inhibited 97.07% and 89.97% respectively by collected purified alpha-amylase inhibitor.