Convenient analytical methods for endo-type glycosidase that acts on glycoconjugates and their application in glycotechnology

Anal Sci. 2003 Jan;19(1):93-7. doi: 10.2116/analsci.19.93.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carbohydrate Sequence
  • Glycoconjugates / chemistry
  • Glycoconjugates / metabolism*
  • Glycoside Hydrolases / analysis*
  • Glycoside Hydrolases / metabolism
  • Molecular Sequence Data
  • Substrate Specificity

Substances

  • Glycoconjugates
  • Glycoside Hydrolases