Synthesis of Phenoxyundecyl Diphosphate Disaccharides for Studies of the Biosynthesis of O Antigenic Polysaccharides in Enteric Bacteria
The biosynthesis of O antigenic polysaccharides in enteric bacteria from nucleoside diphosphate sugars (donor substrates) is catalyzed by the corresponding glycosyltransferases and proceeds through the intermediate formation of undecaprenyl diphosphate sugars (acceptor substrates). To study this process, a chemical synthesis of the compounds having the natural structure or their modified analogs is necessary. The phosphoroimidazolidate method is a universal method for synthesis of lipid diphosphate disaccharides containing 2-acetamido-2-deoxyglycosyl residue at the reducing end of the disaccharide moiety and 11-phenoxyundecyl residue as lipid fragment of the molecule. We report here protocols to synthesize the disaccharides P1-(11-phenoxyundecyl)-P2-(2-acetamido-2-deoxy-3-O-α-d-rhamnopyranosyl-α-d-glucopyranosyl) diphosphate [d-Rha(α1-3)-d-GlcNAcα-PP-PhU, Compound 1] and P1-(11-phenoxyundecyl)-P2-(2-acetamido-2-deoxy-3-O-β-d-galactopyranosyl-α-d-galactopyranosyl) diphosphate [d-Gal(β1-3)-d-GalNAcα-PP-PhU, Compound 6]. We describe the procedures for identification and structure estimation of compounds by TLC, NMR, and MS. We also include the biochemical testing of Compound 6 with α2,3-sialyltransferase WbwA from Escherichia coli O104.
Key wordsChemical synthesis Phosphoroimidazolidate method d-Rha(α1-3)-d-GlcNAcα-PP-PhU d-Gal(β1-3)-d-GalNAcα-PP-PhU TLC NMR MS Sialyltransferase WbwA
This work was supported by the Russian Foundation for Basic Research, Project 16-04-00372 and the Canadian Institutes of Health Research (GlycoNet) (to IB).
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