Separation and direct detection of raw and gelatinized starch hydrolyzing activities of glucoamylase on isoelectric focusing gels.
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 microg protein, it will be useful for enzyme engineering studies that involve screening of various mutations.
Suresh
C
C
Department of Food Microbiology, Central Food Technological Research Institute, Mysore, India.
Dubey
A K
AK
Kini
R
R
Umesh-Kumar
S
S
Karanth
N G
NG
D001234:Aspergillus niger / Q000201:enzymology; D005780:Gelatin / Q000378:metabolism*; D005087:Glucan 1,4-alpha-Glucosidase / Q000378:metabolism*; D006868:Hydrolysis; D007525:Isoelectric Focusing / Q000379:methods*; D013213:Starch / Q000378:metabolism*
D005780:Gelatin; D013213:Starch; D005087:Glucan 1,4-alpha-Glucosidase