Determination of β-Glucosidase Activity Based on Enzyme-Triggered Click Chemistry

Abstract

A novel method to quantify β-glucosidase activity was developed by coupling the cleavage of 2-O-(β-glucopyra-nosyl)ascorbic acid with the reduction of Cu (Ⅱ). The in situ generated Cu (I) catalyzed the cycloaddition between weakly fluorescent coumarin and benzyl azide to yield a highly fluorescent triazole product. The fluorescence intensity was dependently enhanced on the increase of the activity of β-glucosidase and a linear relationship was found between 1~40 U/L. This cascade allows detection of β-glucosidase with a limit of detection of 0.456 U/L.