title: Characterization of recombinant yeast exo-beta-1,3-glucanase (Exg 1p) expressed in Escherichia coli cells.
abstract: Yeast exo-beta-1,3-glucanase gene (EXG1) was expressed in Escherichia coli and the recombinant enzyme (Exg1p) was characterized. The recombinant Exglp had an apparent molecular mass of 45 kDa by SDS-PAGE and the enzyme has a broad specificity for beta-1,3-linkages as well as beta-1,6-linkages, and also for other beta-glucosidic linked substrates, such as cellobiose and pNPG. Kinetic analyses indicate that the enzyme prefers small substrates such as laminaribiose, gentiobiose, and pNPG rather than polysaccharide substrates, such as laminaran or pustulan. With a high concentration of laminaribiose, the enzyme catalyzed transglucosidation forming laminarioligosaccharides. The enzyme was strongly inhibited with high concentrations of laminaran.
authors.affiliation: Divison of Applied Life Science, Graduate School of Agricultural Science, Tohoku University, Sendai, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Suzuki
authors.forename: T
authors.initials: T
authors.lastname: Yabe
authors.forename: Y
authors.initials: Y
authors.lastname: Maruyama
authors.forename: K
authors.initials: K
authors.lastname: Abe
authors.forename: T
authors.initials: T
authors.lastname: Nakajima
mesh_terms: D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology*; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D043326:Glucan 1,3-beta-Glucosidase; D006868:Hydrolysis; D007700:Kinetics; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature; D001617:beta-Glucosidase / Q000032:analysis* / Q000096:biosynthesis / Q000378:metabolism
chemical_list: D003470:Culture Media; D011994:Recombinant Proteins; D001617:beta-Glucosidase; D043326:Glucan 1,3-beta-Glucosidase