title: Purification and characterization of an oligo-isomaltosaccharide synthase from a Streptococcus sobrinus glucosyltransferase-I deficient mutant.
abstract: One glucosyltransferase (GTF) -I deficient mutant of Streptococcus sobrinus strain B13N was isolated through chemical mutagenesis with ethyl methanesulfonate, and characterized. This mutant, designated as B13N-Id, readily allowed us to purify a homogeneous oligo-isomaltosaccharide synthase (GTF-S) from its culture fluid. The purified GTF-S was only recognized with rabbit polyclonal antibody against recombinant GTF-S from an Ecsherichia coli MD124 clone expressing the B13N gtfS gene, and showed the almost same enzymatic properties as the recombinant enzyme. A double reciprocal plot of the B13N GTF-S for sucrose was biphasic, and the affinity for this substrate was high compared to that of GTF-S enzymes from other strains.
authors.affiliation: Department of Microbiology, Nihon University school of Dentistry at Matsudo, Chiba, Japan. nshino@mascat.nihon-u.ac.jp
authors.forename: N
authors.initials: N
authors.lastname: Shinozaki-Kuwahara
authors.forename: M
authors.initials: M
authors.lastname: Hayakawa
authors.forename: T
authors.initials: T
authors.lastname: Shiroza
authors.forename: Y
authors.initials: Y
authors.lastname: Abiko
authors.forename: K
authors.initials: K
authors.lastname: Fukushima
mesh_terms: D002851:Chromatography, High Pressure Liquid; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D005936:Glucans / Q000032:analysis / Q000302:isolation & purification; D005964:Glucosyltransferases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D009154:Mutation; D013291:Streptococcus / Q000201:enzymology* / Q000235:genetics
chemical_list: D003470:Culture Media; D005936:Glucans; D005964:Glucosyltransferases