Capillary zone electrophoresis for the quantitation of oligosaccharides formed through the action of chitinase

Electrophoresis. 1991 Sep;12(9):636-40. doi: 10.1002/elps.1150120907.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amination
  • Calibration
  • Chitinases / metabolism*
  • Electrophoresis / methods
  • Kinetics
  • Magnetic Resonance Spectroscopy / methods
  • Oligosaccharides / analysis*
  • Oxidation-Reduction
  • Protons
  • Spectrometry, Fluorescence

Substances

  • Oligosaccharides
  • Protons
  • Chitinases