Synthesis of highly branched anionic α-glucans by thermostable phosphorylase-catalyzed α-glucuronylation.
Highly branched anionic α-glucans were enzymatically synthesized by thermostable phosphorylase-catalyzed α-glucuronylation of highly branched cyclic dextrin using α-D-glucuronic acid 1-phosphate (GlcA-1-P) as a glycosyl donor. The resulting products were characterized by ¹H NMR measurement as well as high performance anion exchange chromatographic and MALDI-TOF MS analyses after treatments with several amylases. α-D-Glucose 1-phosphate was detected in the reaction mixtures, suggesting the occurrence of phosphorolysis in the α-glucuronylation. The glucuronylation ratios of glucuronic acid residues to non-reducing ends were evaluated from quantification of α-D-glucose 1-phosphate and inorganic phosphate in the reaction mixtures, which were relatively in good agreement with those determined by ¹H NMR analysis of the products. The glucuronylation ratios increased with increasing feed ratios of GlcA-1-P/non-reducing ends.