Purification and characterization of an oligo-isomaltosaccharide synthase from a Streptococcus sobrinus glucosyltransferase-I deficient mutant.
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.
Shinozaki-Kuwahara
N
N
Department of Microbiology, Nihon University school of Dentistry at Matsudo, Chiba, Japan. nshino@mascat.nihon-u.ac.jp
Hayakawa
M
M
Shiroza
T
T
Abiko
Y
Y
Fukushima
K
K
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
D003470:Culture Media; D005936:Glucans; D005964:Glucosyltransferases