β-1,4-galactosyltransferase-catalyzed synthesis of the branched tetrasaccharide repeating unit of Streptococcus pneumoniae type 14

A chemoenzymatic approach is described towards the branched tetrasaccharide repeating unit, beta-D-Galp-(1-->4)-beta-D-Glcp-(1-->6)-[beta-D-Galp-(1-->4)]-beta-D-GlcpNAc, of Streptococcus pneumoniae type 14 in a form suitable for conjugation. The linear trisaccharide acceptor, beta-D-Galp-(1-->4)-beta-D-Glcp-(1-->6)-beta-D-GlcpNAc-(1-->O)CH2CH = CH2, was synthesized by coupling of peracetylated lactosyl trichloroacetimidate to a suitably protected glucosamine building block and subsequent deprotection steps. The obtained derivative was found to be a good acceptor for bovine milk beta-1,4-galactosyltransferase, and the resulting branched tetrasaccharide beta-allyl glycoside was isolated and characterized by NMR spectroscopy and FAB mass spectrometry. Reaction of the anomeric allyl function with cysteamine under UV-irradiation gave the beta-aminoethylthio-extended glycoside suitable for further coupling of the tetrasaccharide to protein carriers. (C) 1998 Elsevier Science Ltd. All rights reserved.