Purification of endo-N-acetyl-beta-D-glucosaminidase H by substrate-affinity chromatography.
摘要：
Endo- N-acetyl-β- d-glucosaminidase H (Endo H) was purified to homogeneity (3000-fold) from a culture filtrate of 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 α- 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 glycopeptide substrate and some protein impurities by gel filtration and hydrophobic chromatography.
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DOI：
10.1016/0008-6215(89)84105-9
被引量：
年份：
1989
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