Purification of endo-N-acetyl-beta-D-glucosaminidase H by substrate-affinity chromatography.
Carbohydr Res
; 189: 289-99, 1989 Jun 15.
Article
em En
| MEDLINE
| ID: mdl-2505925
ABSTRACT
Endo-N-acetyl-beta-D-glucosaminidase H (Endo H) was purified to homogeneity (3000-fold) from a culture filtrate to Streptomyces plicatus. The key step was substrate-affinity chromatography, which afforded a 1000-fold purification and yielded a protease- and exoglycosidase-free preparation of Endo H. Proteins from the crude sample were applied to the substrate-affinity column, consisting of yeast-invertase glycopeptides bound to Sepharose-immobilized concanavalin A. After washing off the unbound proteins, Endo H was quantitatively eluted by methyl alpha-D-mannopyranoside. Various conditions were tested to achieve an optimal binding of Endo H to this substrate-affinity gel. After substrate-affinity chromatography, Endo H was separated from the coeluted gylcopeptide substrate and some protein impurities by gel filtration and hydrophobic chromatography.
Buscar no Google
Base de dados:
MEDLINE
Assunto principal:
Acetilglucosaminidase
/
Hexosaminidases
Idioma:
En
Ano de publicação:
1989
Tipo de documento:
Article