Simultaneous high-performance liquid chromatographic determination of both the cleavage pattern and the stereochemical outcome of the hydrolysis reactions catalyzed by various glycosidases

Abstract A high-performance liquid chromatographic method for the simultaneous determination of both the stereo-chemical outcome and the cleavage pattern of enzymatic action on unmodified sugar substrates is described. Three different enzymes were investigated by this method. Human pancreatic α-amylase hydrolyzed maltopentaose with retention of anomeric configuration, with the cleavage position being two glucose units from the reducing end. Cellulomonas fimi endoglucanase D hydrolyzed cellopentaose with retention of anomeric configuration and predominantly two glucose units from the reducing end. β-D-Xylosidase from Butyrivibrio fibrisolvens hydrolyzed o -nitrophenyl β-D-xylopyranoside with inversion of anomeric configuration.