title: Convenient analytical methods for endo-type glycosidase that acts on glycoconjugates and their application in glycotechnology.
abstract: 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-N-acetylgalactosaminidase 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.
authors.affiliation: Department of Biochemistry, School of Medicine, Kitasato University, 1-15-1 Kitasato, Sagamihara, Kanagawa 228-8555, Japan. ikukara@med.kitasato-u.ac.jp
authors.forename: Ikuko
authors.initials: I
authors.lastname: Ishii-Karakasa
mesh_terms: D000818:Animals; D002240:Carbohydrate Sequence; D006001:Glycoconjugates / Q000737:chemistry / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000032:analysis* / Q000378:metabolism; D008969:Molecular Sequence Data; D013379:Substrate Specificity
chemical_list: D006001:Glycoconjugates; D006026:Glycoside Hydrolases