Enzymatic synthesis of novel disaccharides using disaccharide phosphorylases

A method for the synthesis of novel disaccharides was developed. It involve d the following two steps. The first step consisted of two continuous react ions: the conversion of maltose to beta-D-glucose-1-phosphate and D-glucose by the phosphorolytic activity of maltose phosphorylase and the specific c onsumption of only D-glucose by incubation with glucose-consuming yeast cel ls. The second step involved the addition of an appropriate carbohydrate an d its condensation with the remaining beta-D-glucose-l-phosphate by the syn thetic activity of maltose phosphorylase or trehalose phosphorylase. Severa l factors affecting the yields of disaccharides were optimized. Using this method, five maltose-like derivatives and two trehalose-like derivatives we re synthesized from maltose and the corresponding carbohydrates. Among thes e, 4-O-alpha-D-glucopyranosyl-L-fucopyranose (Glc(alpha 1-4)Fuc) and alpha- D-glucopyranosyl alpha-D-fucopyranoside (Glc(al-la)Fuc) were purified, and identified by H-1 NMR and C-13 NMR.