Fast and reliable method for simultaneous zymographic detection of glucoamylase and α-amylase in fungal fermentation.
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.
Center for Chemistry, Institute of Chemistry, Technology, and Metallurgy, University of Belgrade, Studentski trg 12-16, Belgrade, Serbia.
Biljana
B
Dojnov
Zoran
Z
Vujčić
D001230:Aspergillus / Q000201:enzymology*; D005285:Fermentation; D005087:Glucan 1,4-alpha-Glucosidase / Q000032:analysis*; D015203:Reproducibility of Results; D000516:alpha-Amylases / Q000032:analysis*
D000516:alpha-Amylases; D005087:Glucan 1,4-alpha-Glucosidase