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Separation and direct detection of raw and gelatinized starch hydrolyzing activities of glucoamylase on isoelectric focusing gels
Author(s) -
Suresh Cuddapah,
Dubey Ashok K.,
Kini Ramachandra,
UmeshKumar Sukumaran,
Karanth Naikantatte G.
Publication year - 1999
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(19990301)20:3<483::aid-elps483>3.0.co;2-b
Subject(s) - isoelectric focusing , chemistry , starch , isoelectric point , chromatography , reagent , substrate (aquarium) , biochemistry , aspergillus niger , enzyme , organic chemistry , biology , ecology
A procedure to detect raw and gelatinized starch activities of glucoamylase on isoelectric focusing (IEF) gels by using 2, 3, 5‐triphenyltetrazolium chloride is described. The reagent reacts with the reducing group of glucose released by glucoamylase from the substrate starch. Using the reaction, production of glucoamylase by Aspergillus niger was detected on 10% IEF gels within a pH range of 2.5—9.5. Since the method can detect raw and gelatinized starch activities of glucomylase associated with 1 μg protein, it will be useful for enzyme engineering studies that involve screening of various mutations.