Enzymatic synthesis of novel disaccharides using disaccharide phosphorylases.

Abstract A method for the synthesis of novel disaccharides was developed. It involved the following two steps. The first step consisted of two continuous reactions: the conversion of maltose to β- d -glucose-1-phosphate and d -glucose by the phosphorolytic activity of maltose phosphorylase and the specific consumption of only d -glucose by incubation with glucose-consuming yeast cells. The second step involved the addition of an appropriate carbohydrate and its condensation with the remaining β- d -glucose-1-phosphate by the synthetic activity of maltose phosphorylase or trehalose phosphorylase. Several factors affecting the yields of disaccharides were optimized. Using this method, five maltose-like derivatives and two trehalose-like derivatives were synthesized from maltose and the corresponding carbohydrates. Among these, 4- O -α- d -glucopyranosyl- l -fucopyranose (Glc(α1–4)Fuc) and α- d -glucopyranosyl α- d -fucopyranoside (Glc(α1-1α)Fuc) were purified, and identified by 1 H NMR and 13 C NMR.