Synthesis of L-Rhamnose and N -Acetyl-D-Glucosamine Derivatives Entering in the Composition of Bacterial Polysaccharides by Use of Glucansucrases

Abstract

Transglucosylation reactions using sucrose as glucosyl donor and either N-acetyl-D-glucosamine, L-rhamnose, or methyl α -L-rhamnopyranoside as acceptors were carried out with recombinant glucansucrases from families 70 and 13 of glycoside-hydrolases. Depending on the enzyme specificity, various carbohydrate structures were synthesized and characterized including α -D-glucopyranosyl-(1 → 6)-N-acetyl-D-glucosamine, α -D-glucopyranosyl-(1 → 4)-N-acetyl-D-glucosamine, α -D-glucopyranosyl-(1 → 1)-β -L-rhamnopyranoside, α -D-glucopyranosyl-(1 → 4)-α -D-glucopyranosyl-(1 → 1)-β -L-rhamnopyranoside, methyl α -D-glucopyranosyl-(1 → 4)-α -L-rhamnopyranoside, and methyl α -D-glucopyranosyl-(1 → 3)-α -L-rhamnopyranoside. Disaccharides were obtained with yields going up to 64%. The structural diversity generated as well as the obtained yields appear to be related to enzyme active site architecture, which can be modulated and improved by enzyme engineering. Several of the obtained disaccharides enter in the composition of surface polysaccharides of pathogenic bacteria, among which is Shigella flexneri. Our results outline the potential of glucansucrases in the chemo-enzymatic synthesis of complex carbohydrates of biological interest whose chemical synthesis may be seen as a limitation.