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

Anal Biochem

Center for Chemistry, Institute of Chemistry, Technology, and Metallurgy, University of Belgrade, Studentski trg 12-16, Belgrade, Serbia.

Published: February 2012

Detection of α-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. α-Amylase was detected on the substrate gel with copolymerized β-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.

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http://dx.doi.org/10.1016/j.ab.2011.11.039DOI Listing

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