Determination of alpha-amylase using a new blocked substrate (3-ketobutylidene beta-2-chloro-4-nitrophenyl-maltopentaoside).
A new substrate, 3-ketobutylidene beta-2-chloro-4-nitrophenylmaltopentaoside (3KB-CNPG5), was used for the determination of alpha-amylase (EC 3.2.1.1) in serum and urine. Under this alpha-amylase assay condition, 3KB-CNPG5 is resistant to glucoamylase and alpha-glucosidase, which are auxiliary enzymes, because the 4- and 6-positions of the non-reducing-end glucose residue are modified by the 3-ketobutylidene group. The assay using 3KB-CNPG5 for alpha-amylase activity is a highly sensitive method that uses 2-chloro-4-nitrophenol (CNP) as an aglycone, and is a stable method for determination of alpha-amylase activity in biological fluids.
Tsuruga Enzyme Plant, Toyobo Co. Ltd., Fukui and Japan.
S
S
Teshima
Y
Y
Hayashi
S
S
Emi
K
K
Ishimaru
D002851:Chromatography, High Pressure Liquid; D005959:Glucosidases / Q000378:metabolism; D005960:Glucosides; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D007700:Kinetics; D010179:Pancreas / Q000201:enzymology; D012463:Saliva / Q000201:enzymology; D013056:Spectrophotometry, Ultraviolet; D013379:Substrate Specificity; D000516:alpha-Amylases / Q000032:analysis* / Q000037:antagonists & inhibitors
D005960:Glucosides; D007202:Indicators and Reagents; C071715:3-ketobutylidene 2-chloro-4-nitrophenylmaltopentaoside; D005959:Glucosidases; D000516:alpha-Amylases