Analytical Sciences

Convenient Analytical Methods for Endo-type Glycosidase that Acts on Glycoconjugates and Their Application in Glycotechnology

Ikuko ISHII-KARAKASA

Department of Biochemistry, School of Medicine, Kitasato University, 1-15-1 Kitasato, Sagamihara, Kanagawa 228-8555, Japan

The following procedures were established in order to develop useful degradation enzymes of glycoconjugate for developing postgenome and postproteome research: (1) Enzyme activity with a short time reliability was measured using small amounts by HPLC. (2) The structures of the sugar chains liberated from the glycoconjugate were non-destructively analyzed using small amounts of sugar chains only by 1D ¹H-NMR and H-H COSY spectrometry and a computer simulation of the spectrum. (3) The conformations of the sugar chains liberated from a glycoconjugate in aqueous solution were estimated using 1D ¹H-NMR and H-H COSY spectrometry and the anisotropic effect. Endo-alpha-Nacetylgalactosaminidase from the culture medium of Streptomyces sp. OH-11242 developed using the above methods transferred the sugar chain to sugars and peptides; therefore, it was also an effective enzyme when synthesizing sugar chains and glycopeptides.

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