Automatic spectrophotometric determination of amyloglucosidase activity using p-nitrophenyl-α-D-glucopyranoside and a flow injection analyser

Abstract

An automated flow injection analysis (FIA) method has been developed, using as a chromophore p-nitrophenyl-α-D-glucopyranoside, which is hydrolysed to glucose and p-nitrophenol by amyloglucosidase. The p-nitrophenol is determined spectrophotometrically at 400 nm as a measure of the enzyme activity. The general conditions of the analysis have been optimised. The analytical method is flexibile, as all methods for the analysis of enzymes using soluble p-nitrophenyl derivatives as chromophores may in principle be carried out with the FIA method described. The final result is a simple, automated method with high precision, sensitivity and speed.