Many oligosaccharides are not commercially available, which limits studies focused on elucidation of glycan functions; therefore chemo-enzymatic methods to synthesize them can be very useful. Here, we describe the procedure to synthesize the Galα1-3GalNAcβ1-4GlcNAcβ-R (Gal-LDN) moiety, containing the Galα1-3GalNAc epitope found on the parasitic helminth Haemonchus contortus. An acceptor substrate providing a terminal N-acetylglucosamine was prepared by coupling the fluorescent hydrophobic aglycon, 2,6-diaminopyridine (DAP), to N,N′-diacetylchitobiose. By the subsequent action of recombinant Caenorhabditis elegans β1,4-N- acetylgalactosaminyltransferase the substrate was efficiently converted to GalNAcβ1-4GlcNAcβ-R (LDN-R). Since no recombinant α1,3- galactosyltransferase has been described that acts on terminal βGalNAc, we used bovine α1,3-galactosyltransferase to obtain a partial conversion of LDN-R to the Gal-LDN antigen. This method can be applied to synthesize any oligosaccharide, provided that specific glycosyltransferases are available, or related enzymes that can be pushed to elongate the selected acceptor. © Springer Science+Business Media New York 2013.
CITATION STYLE
Tefsen, B., & Van Die, I. (2013). Glycosyltransferases in chemo-enzymatic synthesis of oligosaccharides. Methods in Molecular Biology, 1022, 357–367. https://doi.org/10.1007/978-1-62703-465-4_27
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