Abstract
Cellobiose phosphorylase from Cellvibrio gilvus was examined for its acceptor specificity in the synthetic reaction with glucose-1-phosphate, using substrates in which the C-6 substituent of d-Glc had been altered. A range of disaccharides were also tested for acceptor specificity but only those with (1⃗6)-linkages were successful acceptors. Melibiose, gentiobiose, isomaltose and also the monosaccharide glucuronamide were found to react with cellobiose phosphorylase and glucose-1-phosphate giving β- d-Glc p-(1⃗4)-[ α- d-Gal p-(1⃗6)]- d-Glc p, β- d-Glc p-(1⃗4)-[ β- d-Glc p-(1⃗6)]- d-Glc p, β- d-Glc p-(1⃗4)-[ α- d-Glc p-(1⃗6)]- d-Glc p and β- d-Glc p-(1⃗4)- d-GlcUN p, respectively. These products were purified using a range of chromatographic methods and characterised by NMR and FAB-MS. This is the first time cellobiose phosphorylase has been shown to synthesise trisaccharides.
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