Simultaneous high-performance liquid chromatographic determination of both the cleavage pattern and the stereochemical outcome of the hydrolysis reactions catalyzed by various glycosidases.
A high-performance liquid chromatographic method for the simultaneous determination of both the stereochemical outcome and the cleavage pattern of enzymatic action on unmodified sugar substrates is described. Three different enzymes were investigated by this method. Human pancreatic alpha-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. beta-D-Xylosidase from Butyrivibrio fibrisolvens hydrolyzed o-nitrophenyl beta-D-xylopyranoside with inversion of anomeric configuration.
Braun
C
C
Department of Chemistry, University of British Columbia, Vancouver, Canada.
Meinke
A
A
Ziser
L
L
Withers
S G
SG
D000192:Actinomycetales / Q000201:enzymology; D001665:Binding Sites; D002480:Cellulase / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006089:Gram-Negative Anaerobic Bacteria / Q000201:enzymology; D006801:Humans; D006868:Hydrolysis; D009844:Oligosaccharides / Q000737:chemistry; D010179:Pancreas / Q000201:enzymology; D013237:Stereoisomerism; D013379:Substrate Specificity; D014995:Xylosidases / Q000378:metabolism; D000516:alpha-Amylases / Q000378:metabolism
D009844:Oligosaccharides; D006026:Glycoside Hydrolases; D014995:Xylosidases; D000516:alpha-Amylases; D002480:Cellulase