Alpha-amylase inhibitors found in seeds are drug-design targets for treatment of diabetes and digestion disorders. These inhibitors are also known as sensitizing agents in humans. Plant alpha-amylase inhibitors show great potential as tools to engineer resistance of crop plants against pests. Numerous forms of alpha –amylase inhibitors have been reported. Alphaamylase 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 a 0.59 relative mobility. Total hydrolytic activity of human salivary and bacillus subtilis alpha –amylase were inhibited 92.01% and 78.99%, respectively by purified alpha- amylase inhibitor.