Fast and reliable method for simultaneous zymographic detection of glucoamylase and a-amylase in fungal fermentation

Abstract

Detection of alpha-amylase and glucoamylase in crude fermentation extracts using a single native electrophoresis gel and zymogram is described in this article. Proteins were printed on substrate gel and simultaneously onto a membrane in a three-sandwich gel. alpha-Amylase was detected on the substrate gel with copolymerized beta-limit dextrins and iodine reagent. Glucoamylases were detected on the membrane using a coupled assay for glucose detection. Both amylases were detected in native gel using starch and iodine reagent. The described technique can be a helpful tool for monitoring and control of fermentation processes because fungal amylase producers almost always synthesize both amylases. (C) 2011 Elsevier Inc. All rights reserved.