Structure and biosynthetic locus of the lipopolysaccharide outer core produced by Pasteurella multocida serovars 8 and 13 and the identification of a novel phospho-glycero moiety

Pasteurella multocida strains are classified into 16 Heddleston serovars on the basis of the lipopolysaccharide (LPS) antigens expressed on the surface of the bacteria. The LPS structure and the corresponding LPS outer core biosynthesis loci of strains belonging to serovars 1, 2, 3, 5, 9 and 14 have been characterized, revealing a clear structural basis for serovar classification. However, several of these serovars are genetically related, sharing the same LPS outer core biosynthesis locus, but producing different LPS molecules as a result of mutations within LPS assembly genes. In this article, we report that the P. multocida type strains belonging to serovars 8 and 13 share the same LPS outer core biosynthesis locus and produce structurally related LPS molecules. Structural analysis of the serovar 8 LPS revealed an inner core that is conserved among P. multocida strains and the following outer core structure: where X is a unique phospho-glycero moiety, 1-((4-aminobutyl)amino)-3-hydroxy-1-oxopropan-2-yl hydrogen phosphate, attached to the sixth position of (1S)GalaNAc. For serovar 13, the LPS structure is the same except for the absence of the terminal phospho-glycero moiety. Analysis of the common outer core biosynthesis locus from the serovar 8 and 13 type strains identified three genes that we predict are involved in the biosynthesis of this terminal moiety. Furthermore, bioinformatic comparisons with the characterized LPS outer core glycosyltransferases from Actinobacillus pleuropneumoniae serovar 1, strain 4074, allowed us to assign a function for each of the glycosyltransferases encoded within the serovar 8/13 LPS outer core biosynthesis locus.