Synthesis and structural analysis of five novel oligosaccharides prepared by glucosyltransfer from beta-D-glucose 1-phosphate to isokestose and nystose using Thermoanaerobacter brockii kojibiose phosphorylase.
Five novel oligosaccharides (tetra-, penta- and hexa-saccharides) were synthesized by glucosyltransfer from beta-D-glucose 1-phosphate to isokestose (O-beta-D-fructofuranosyl-(2-->1)-O-beta-D-fructofuranosyl-(2-->1)-alpha-D-glucopyranoside) or nystose (O-beta-D-fructofuranosyl-(2-->1)-O-beta-D-fructofuranosyl-(2-->1)-O-beta-D-fructofuranosyl-(2-->1)-alpha-D-glucopyranoside) using Thermoanaerobacter brockii kojibiose phosphorylase. The oligosaccharides were identified as 2(2-alpha-D-glucopyranosyl)(m)isokestose; [O-alpha-D-glucopyranosyl-(1-->2)](m)-O-[beta-D-fructofuranosyl-(2-->1)](2)-alpha-D-glucopyranoside: m=1, 2, and 3, and 2(2-alpha-D-glucopyranosyl)(n)nystose; [O-alpha-D-glucopyranosyl-(1-->2)](n)-O-[beta-D-fructofuranosyl-(2-->1)](3)-alpha-D-glucopyranoside: n=1 and 2 using gas liquid chromatography analysis of the methyl derivatives, and MALDI-TOF-MS and NMR measurements of the newly formed oligosaccharides. 1H, 13C NMR signals of each saccharide were assigned using 2D-NMR techniques, including COSY, HSQC, HSQC-TOCSY, HMBC, CH(2)-selected E-HSQC, and CH(2)-selected E-HSQC-TOCSY.
Okada
Hideki
H
Department of Food Production and Utility Development, Graduate School of Dairy Science Research, Rakuno Gakuen University, Ebetsu 069-8501, Japan.
Fukushi
Eri
E
Onodera
Shuichi
S
Nishimoto
Tomoyuki
T
Kawabata
Jun
J
Kikuchi
Masanori
M
Shiomi
Norio
N
D001421:Bacteria, Anaerobic / Q000201:enzymology*; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D004187:Disaccharides / Q000378:metabolism*; D005958:Glucosephosphates / Q000737:chemistry*; D006358:Hot Temperature; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000138:chemical synthesis* / Q000737:chemistry* / Q000302:isolation & purification; D006005:Phosphorylases / Q000378:metabolism*; D014312:Trisaccharides / Q000737:chemistry*
D004187:Disaccharides; D005958:Glucosephosphates; D009844:Oligosaccharides; D014312:Trisaccharides; C043600:fungitetraose; C043601:isokestose; C008237:kojibiose; C031590:glucose-1-phosphate; D006005:Phosphorylases