title: Capillary zone electrophoresis for the quantitation of oligosaccharides formed through the action of chitinase.
abstract: Capillary zone electrophoresis with fluorescence detection was used to analyze the products formed by chitinase acting on N-acetylchitooligosaccharide-fluorescent conjugates. Six oligosaccharides of the structure [N-acetylglucosamine(1----4)]n (where n = 1-6) were conjugated to 7-amino-1,3-naphthalene disulfonic acid by reductive amination. Each oligosaccharide-fluorescent conjugate was purified by preparative gradient polyacrylamide gel electrophoresis, semi-dry electrotransfer to a positively-charged nylon membrane and recovered by washing the membrane with salt solution. The products formed by treating each oligosaccharide-fluorescent conjugate with chitinase were analyzed by capillary zone electrophoresis. The chitinase treatment hexasaccharide-fluorescent conjugate was also examined kinetically to study the action pattern of this enzyme.
authors.affiliation: Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City 52242.
authors.forename: K B
authors.initials: KB
authors.lastname: Lee
authors.forename: Y S
authors.initials: YS
authors.lastname: Kim
authors.forename: R J
authors.initials: RJ
authors.lastname: Linhardt
mesh_terms: D000586:Amination; D002138:Calibration; D002688:Chitinases / Q000378:metabolism*; D004586:Electrophoresis / Q000379:methods; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D009844:Oligosaccharides / Q000032:analysis*; D010084:Oxidation-Reduction; D011522:Protons; D013050:Spectrometry, Fluorescence
chemical_list: D009844:Oligosaccharides; D011522:Protons; D002688:Chitinases