Determination of α-amylase using a new blocked substrate (3-ketobutylidene β-2-chloro-4-nitrophenyl-maltopentaoside)

• Published in: Clinica chimica acta Vol. 199; no. 1; pp. 23 - 31
• Main Authors: Teshima, Shinichi, Hayashi, Yuzo, Emi, Shigenori, Ishimaru, Katsutosi
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 31.05.1991
• Subjects: 3-Ketobutylidene β-2-chloro-4-nitrophenyl-maltopentaoside; alpha-Amylases - analysis; alpha-Amylases - antagonists & inhibitors; Chromatography, High Pressure Liquid; Glucosidases - metabolism; Glucosides; Humans; Hydrogen-Ion Concentration; Indicators and Reagents; Kinetics; Pancreas - enzymology; Saliva - enzymology; Spectrophotometry, Ultraviolet; Substrate Specificity; α-Amylase; 3-Ketobutylidene β-2-chloro-4-nitrophenyl-maltopentaoside; α-Amylase
• ISSN: 0009-8981; 1873-3492
• DOI: 10.1016/0009-8981(91)90005-W

A new substrate, 3-ketobutylidene β-2-chloro-4-nitrophenylmaltopentaoside (3KB-CNPG5), was used for the determination of α-amylase (EC 3.2.1.1) in serum and urine. Under this α-amylase assay condition, 3KB-CNPG5 is resistant to glucoamylase and α-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 α-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 α-amylase activity in biological fluids.