A spectrophotometric assay for glucosidase I.
A spectrophotometric assay for glucosidase I using the synthetic trisaccharide alpha-D-Glc 1-->2 alpha-D-Glc 1-->3 alpha-D-Glc-O(CH2)8COOCH3 is reported. The terminal glucose is released from the substrate by the enzyme and quantitated using glucose oxidase, peroxidase, and o-dianisidine. The trisaccharide is specific for glucosidase I and provides all the necessary structural features for correct interaction in the enzyme active site. The utility of the assay for monitoring enzyme activity during isolation and for use in kinetic and inhibition studies (i.e., with 1-deoxynorjirimycin) is demonstrated.
Department of Chemistry, University of Alberta, Edmonton, Canada.
I
I
Neverova
C H
CH
Scaman
O P
OP
Srivastava
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R
Szweda
I K
IK
Vijay
M M
MM
Palcic
D017485:1-Deoxynojirimycin / Q000494:pharmacology; D002240:Carbohydrate Sequence; D002852:Chromatography, Ion Exchange; D065089:Glycoside Hydrolase Inhibitors; D007700:Kinetics; D008861:Microsomes / Q000201:enzymology; D008969:Molecular Sequence Data; D013053:Spectrophotometry / Q000379:methods*; D000520:alpha-Glucosidases / Q000032:analysis*
D065089:Glycoside Hydrolase Inhibitors; D017485:1-Deoxynojirimycin; C042059:glucosidase I; D000520:alpha-Glucosidases