A Unique Sugar l-Perosamine (4-Amino-4,6-dideoxy-l-mannose) Is a Compound Building Two O-Chain Polysaccharides in the Lipopolysaccharide of Strain JCM 3968, Serogroup O6.

Lipopolysaccharide (LPS) is the major glycolipid and virulence factor of Gram-negative bacteria, including spp. The O-specific polysaccharide (O-PS, O-chain, O-antigen), i.e., the surface-exposed part of LPS, which is a hetero- or homopolysaccharide, determines the serospecificity of bacterial strains. Here, chemical analyses, mass spectrometry, and H and C NMR spectroscopy techniques were employed to study the O-PS of strain JCM 3968, serogroup O6. MALDI-TOF mass spectrometry revealed that the LPS of JCM 3968 has a hexaacylated lipid A with conserved architecture of the backbone and a core oligosaccharide composed of HepHexHexNHexNAcKdoP. To liberate the O-antigen, LPS was subjected to mild acid hydrolysis followed by gel-permeation-chromatography and revealed two O-polysaccharides that were found to contain a unique sugar 4-amino-4,6-dideoxy-l-mannose (-acetyl-l-perosamine, l-Rha4NAc), which may further determine the specificity of the serogroup. The first O-polysaccharide (O-PS1) was built up of trisaccharide repeating units composed of one α-d-GalNAc and two α-l-Rha4NAc residues, whereas the other one, O-PS2, is an α1→2 linked homopolymer of l-Rha4NAc. The following structures of the O-polysaccharides were established: O-PS1 →3)-α-l-Rha4NAc-(1→4)-α-d-GalNAc-(1→3)-α-l-Rha4NAc-(1→ O-PS2 →2)-α-l-Rha4NAc-(1→ The present paper is the first work that reveals the occurrence of perosamine in the l-configuration as a component of bacterial O-chain polysaccharides.