Abstract
A method for detection of electrophoretically separated proteinaceous amylase inhibitors is described. Seed powders of pigeon pea, sorghum, chick pea and pearl millet were extracted with 0.1 M HCl and the amylase inhibitors present in the extract were analyzed after ammonium sulfate fractionation. The inhibitors were separated in polyacrylamide gel containing 0.5% soluble starch by electrophoresis and visualized by incubation of the gel in salivary amylase solution and staining with iodine. Starch in the gel is hydrolyzed by amylase during incubation, but the starch in vicinity of amylase inhibitor is protected from hydrolysis and appears as a blue band after staining. Using this protocol, pigeonpea, sorghum, chick pea and pearl millet seed extracts were found to contain at least three inhibitors of salivary amylase. These inhibitors had no activity against bacterial and fungal amylases. The method can be used to screen specificity of individual amylase inhibitors against various amylases.
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