Glucosyl-L-Sorbose, -maltose, and -sucrose were synthesized using kojibiose phosphorylase (KPase) from Thermoanaerobacter brockii ATCC35047 with beta -D-glucose-1-phosphate (beta -G1P) as a glucosyl donor. One disaccharide and two trisaccharides thus synthesized were isolated by Toyopearl HW-40S column chromatography. The results of KPase digestion, methylation analysis, and C-13-NMR studies indicated that these oligosaccharides were alpha -L-glucopyranosyl-(1 -->2)-alpha -L-sorbopyranose, alpha -D-glucopyranosyl-(1 -->2)-alpha -D-glucopyranosyl-(1 -->4)-D-glucopyranose (4-alpha -D-kojibiosyl-glucose), and alpha -D-glucopyranosyl-(1 -->2)-alpha -D-glucopyranosyl-(1 -->2)-alpha -D-fructofuranoside, which are all novel oligosaccharides. Glucosyl-L-sorbose was partially hydrolyzed to glucose and L-sorbose by alpha -glucosidases, while glucosyl-sucrose and glucosyl-maltose were not hydrolyzed by glucoamylase, alpha -glucosidases, or CGTase.