title: Purification of the oligosaccharide-cleaving enzymes of Flavobacterium meningosepticum.
abstract: Four oligosaccharide chain-cleaving enzymes, including two new endoglycosidases distinct from endo-beta-acetylglucosaminidase (Endo) F1, have been identified and purified to homogeneity from cultural filtrates of Flavobacterium meningosepticum. FPLC-directed hydrophobic-interaction chromatography in conjunction with high-resolution ion-exchange chromatography provided a more simple, rapid method for the isolation of endoglycosidase F1, F2 and F3, and the amidase, peptide-N4-N-acetyl-beta-D-glucosaminyl)-asparagine amidase (PNGase F), in greater than 50% yield. The specificity of PNGase F and Endo F1 are well established. Endo F2 and Endo F3 represent new distinct endoglycosidases that prefer complex as compared to high-mannose asparagine-linked glycans. Endo F2 cleaved biantennary oligosaccharides, whereas Endo F3 cleaved both bi- and triantennary oligosaccharides.
authors.affiliation: Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany 12201-0509.
authors.forename: T H
authors.initials: TH
authors.lastname: Plummer
authors.forename: A L
authors.initials: AL
authors.lastname: Tarentino
mesh_terms: D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D005417:Flavobacterium / Q000201:enzymology*; D006026:Glycoside Hydrolases / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D009844:Oligosaccharides / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D009844:Oligosaccharides; D006026:Glycoside Hydrolases