title: Purification and characterization of membrane-bound endoglycoceramidase from Corynebacterium sp.
abstract: Endoglycoceramidase catalyzes the hydrolysis of the linkage between oligosaccharides and ceramides of various glycosphingolipids. We found that a bacterial strain Corynebacterium sp., isolated from soil, produced endoglycoceramidase both intracellularly and extracellularly. The intracellular enzyme bound to the cell membrane was solubilized with 1% Triton X-100 and purified to homogeneity about 170-fold with 60% recovery. The molecular mass of the enzyme was approximately 65 kDa. The enzyme is most active at pH 5.5-6.5 and stable at pH 3.5-8.0. Various neutral and acidic glycosphingolipids were hydrolyzed by the enzyme in the presence of 0.1% Triton X-100. Ganglio- and lacto-type glycosphingolipids were readily hydrolyzed, but globo-type glycosphingolipids were hydrolyzed slowly.
authors.affiliation: Research Laboratory of Higashimaru Shoyu Co. Ltd, Hyogo, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Ashida
authors.forename: K
authors.initials: K
authors.lastname: Yamamoto
authors.forename: H
authors.initials: H
authors.lastname: Kumagai
authors.forename: T
authors.initials: T
authors.lastname: Tochikura
mesh_terms: D002240:Carbohydrate Sequence; D002413:Cations, Divalent; D002462:Cell Membrane / Q000201:enzymology; D002845:Chromatography; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D003352:Corynebacterium / Q000201:enzymology*; D003902:Detergents / Q000494:pharmacology; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D006026:Glycoside Hydrolases / Q000302:isolation & purification* / Q000378:metabolism*; D006028:Glycosphingolipids / Q000378:metabolism*; D006882:Hydroxyapatites; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016339:Spectrometry, Mass, Fast Atom Bombardment; D013379:Substrate Specificity
chemical_list: D002413:Cations, Divalent; D003902:Detergents; D006028:Glycosphingolipids; D006882:Hydroxyapatites; D017886:Durapatite; D006026:Glycoside Hydrolases; C050427:endoglycoceramidase