Structural characterization of two oligosaccharide fragments formed by the selective cleavage of rhamnogalacturonan II: evidence for the anomeric configuration and attachment sites of apiose and 3-deoxy-2-heptulosaric acid

Abstract

Evidence for the anomeric configurations and attachment sites of 3-deoxy-D-lyxo-2-heptulosaric acid (DHA) and apiosyl residues has been obtained through the characterization of two oligoglycosyl fragments isolated from rhamnogalacturonan II (RG-II). One of the oligoglycosyl fragments, a pentaglycosyl aldonic acid generated by Smith degradation of RG-II, was composed of four D-galactopyranosyluronic acid residues, a DHA residue, and a threonic acid residue (derived from a D-galactopyranosyluronic acid residue). The structural analysis of the pentaglycosyl aldonic acid established the beta-D-configuration for the DHA residue. Furthermore, it established that a previously identified diglycosyl side chain, 5-O-(beta-L-arabinofuranosyl)-DHA was directly attached to O-3 of a D-galactopyranosyluronic acid residue in the backbone of RG-II. The second oligoglycosyl fragment, a peralkylated diglycosyl hex-1-enitol, was generated by hex-5-enose degradation of permethylated and carboxyl-reduced RG-II. The structure of the peralkylated diglycosyl hex-1-enitol, beta-L-Rhap-(1----3')-beta-D-Apif-(1----5)-hex-1-enitol++ +, was determined by a combination of glycosyl-linkage composition analysis and n.m.r. spectroscopy. The n.m.r. data indicated the beta-configuration for the D-apiosyl residue. The isolation and characterization of the diglycosyl hex-1-enitol also established that a previously identified heptaglycosyl side chain was directly attached to O-2 of a D-galactopyranosyluronic acid in the backbone of RG-II.