Efficient one-pot enzymatic synthesis of a-(1?4)-glucosidic disaccharides through a coupled reaction catalysed by Lactobacillus acidophilus NCFM maltose phosphorylase

摘要

Lactobacillus acidophilus NCFM maltose phosphorylase (LaMalP) of glycoside hydrolase family 65 catalysed enzymatic synthesis of a-(1?4)-glucosidic disaccharides from maltose and five monosaccharides in a coupled phosphorolysis/reverse phosphorolysis one-pot reaction. Thus phosphorolysis of maltose to b-glucose 1-phosphate circumvented addition of costly b-glucose 1-phosphate for reverse phosphorolysis with different monosaccharide acceptors, resulting in 91%, 89%, 88%, 86% and 84% yield of a-D-glucopyranosyl-(1?4)-N-acetyl-D-glucosaminopyranose [N-acetyl-maltosamine], a-D-glucopyranosyl- (1?4)-D-glucosaminopyranose [maltosamine], a-D-glucopyranosyl-(1?4)-D-mannopyranose, a-D-glucopyranosyl-(1?4)-L-fucopyranose and a-D-glucopyranosyl-(1?4)-D-xylopyranose, respectively, from 0.1 M maltose, 0.5 M N-acetyl glucosamine, 0.1 M glucosamine, 0.1 M mannose, 1 M L-fucose and 0.5 M xylose in 0.2 M phosphate-citrate pH 6.2. These current yields of 0.27–0.34 g of disaccharide products from 10 mL reaction mixtures are easy to scale up and moreover the strategy can be applied to large-scale production of other oligosaccharides from low-cost disaccharides as catalysed by phosphorylases with different substrate specificities.