Colorimetric Assay for Evaluating Glycosyl Fluoride-hydrolyzing Activity of Glycosidase by Using Alizarin Complexon Reagent

摘要

A novel enzyme assay for screening glycosidases has been developed by using glycosyl fluorides.The method is based on the color change caused by the complex formation of fluoride ion and lanthanum-alizarin complexon (La3+-ALC).The assay showed higher sensitivity compared with the conventional method using p-nitrophenyl glycoside as screening substrate.A quantitative study on the hydrolysis of jS-lactosyl fluoride catalyzed by a purified celhilase has been demonstrated,indicating that the present method can also be employed for enzyme kinetics.