Structure of the oligosaccharide chain of the SR-type lipopolysaccharide of Ralstonia solanacearum Toudk-2

Aiming at improving classification and taxonomy of Gram-negative phytopathogenic bacteria, we studied the structure of the lipopolysaccharide of Ralstonia solanacearum. Mild acid hydrolysis of the lipopolysaccharide of strain Toudk-2 followed by gel chromatography resulted in an O-polysaccharide and two oligosaccharide fractions. The smallest-size oligosaccharide fraction was studied by sugar analysis, high-resolution electrospray ionization mass spectrometry, and, after fractionation by anion-exchange chromatography on HiTrap Q, by one- and two-dimensional (1)H and (13)C NMR spectroscopy. It was found that the isolated oligosaccharides consist of the lipopolysaccharide core with one O-polysaccharide repeat (O-unit) attached. The core exists in two major glycoforms differing from each other in a lateral octulosonic acid residue, which is either D-glycero-D-talo-oct-2-ulosonic acid or 3-deoxy-D-manno-oct-2-ulosonic acid. A peculiar feature of the core is the occurrence of 4-amino-4-deoxy-L-arabinose nonstoichiometrically linked to a heptose residue. The full structures of the core and the biological O-unit as well as the site of the attachment of the O-unit to the core were established.