title: Purification and characterization of beta-1,6-glucanase of Streptomyces rochei application in the study of yeast cell wall proteins.
abstract: A beta-1,6-glucanase was purified to apparent homogeneity from a commercial yeast digestive enzyme prepared from Streptomyces rochei by a series of column chromatographies. The molecular mass of the purified enzyme was 60 kDa by SDS-PAGE. The purified enzyme had an optimum pH range from 4.0 to 6.0 and was stable in the same pH range. The enzyme was stable under 50 degrees C but lost almost all activity at 60 degrees C. The enzyme was specific to beta-1,6-glucan and had little activity towards beta-1,3-glucan and beta-1,4-glucan. When the beta-1,6-glucan was hydrolyzed with the purified enzyme for 5 h, the reaction products contained 20% glucose, 36% gentiobiose, and 44% other oligosaccharides, suggesting that the enzyme is an endo-type glucanase. When the purified enzyme was used for the digestion of the cell wall of Saccharomyces cerevisiae, cell-wall proteins covalently bound to the cell-wall glucan were recovered as soluble forms, suggesting that this enzyme is useful for analysis of yeast-cell wall proteins.
authors.affiliation: National Research Institute of Brewing, 3-7-1 Kagamiyama, Higashihiroshima 739-0046, Japan. wuhong@nrib.go.jp
authors.forename: Hong
authors.initials: H
authors.lastname: Wu
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Shimoi
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Ito
mesh_terms: D002473:Cell Wall / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000378:metabolism*; D005936:Glucans / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D013302:Streptomyces / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D005656:Fungal Proteins; D005936:Glucans; D006026:Glycoside Hydrolases; C020105:endo-1,6-beta-glucanase