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Capillary zone electrophoresis for the quantitation of oligosaccharides formed through the action of chitinase
Author(s) -
Lee KyungBok,
Kim YeongShik,
Linhardt Robert J.
Publication year - 1991
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150120907
Subject(s) - capillary electrophoresis , chemistry , chromatography , conjugate , fluorescence , oligosaccharide , chitinase , reductive amination , electrophoresis , polyacrylamide gel electrophoresis , gel electrophoresis , biochemistry , enzyme , mathematical analysis , physics , mathematics , quantum mechanics , catalysis
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.