Electrophoretic transfer as a technique for the detection and identification of plant amylolytic enzymes in polyacrylamide gels

Abstract

An electrophoretic transfer technique was developed for the specific identification of isozymes of starch debranching enzyme, alpha-amylase, and beta-amylase. Amylolytic enzymes are separated by native polyacrylamide slab gel electrophoresis and proteins in gels are electrophoretically transferred through starch-containing polyacrylamide gels. Each amylolytic enzyme degrades starch in the transfer gel to its characteristic limit dextrin as it moves through the gel. Various limit dextrins in the starch gel are identified by their characteristic color development in KI/I solution. Isozymes of starch debranching enzyme, alpha-amylase, and beta-amylase can be easily identified in the same gel.