A series of (LacNAc)n oligomers with n=1-4 (at first with [13C] in natural abundance then enriched in [13C]-Gal and/or [13C]-GlcNAc at specific sites in the sequence) are being prepared by enzymatic synthesis using the appropriate glycosyltransferases and sugar nucleotide substrates. Specifically, oligo-N-acetyllactosamine synthesis is being performed using human milk b(1,4)-galactosyltransferase and a system of coupled reactions that produce UDP-Gal (the sugar nucleotide substrate for this enzyme) as an intermediate in the reaction. The Pierce laboratory has purified b(1,3)-N-acetylglucosaminyltransferase (GnT-I, from human serum), the enzyme needed to elongate lactose/lactosamine. To date, hexamers of both the lacto-neotetraose series as well as the lactosamine series have been synthesized and their structures confirmed by NMR spectroscopy. Preliminary binding studies on the lacto-neotetraose series have been completed using line shape analysis of 1H-NMR spectra.
Showing the most recent 10 out of 245 publications
