Journal
CARBOHYDRATE RESEARCH
Volume 345, Issue 17, Pages 2523-2528Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.carres.2010.09.007
Keywords
Outer-core Kdo; NMR spectroscopy; Plesiomonas; Reductive hydrolysis; Structure elucidation
Funding
- Ministerio de Educacion, Ciencia y Deporte and Ministerio de Sanidad, Spain
- Generalitat de Catalunya (Centre de Referencia en Biotecnologia)
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Plesiomonas shigelloides is a Gram-negative opportunistic pathogen associated with gastrointestinal and extraintestinal infections, which especially invades immunocompromised patients and neonates. The lipopolysaccharides are one of the major virulence determinants in Gram-negative bacteria and are structurally composed of three different domains: the lipid A. the core oligosaccharide and the O-antigen polysaccharide. In the last few years we elucidated the structures of the O-chain and the core oligosaccharide from the P. shigelloides strain 302-73. In this paper we now report the characterization of the linkage between the core and the O-chain. The LPS obtained after PCP extraction contained a small number of O-chain repeating units. The product obtained by hydrazinolysis was analysed by FTICR-ESIMS and suggested the presence of an additional Kdo in the core oligosaccharide. Furthermore, the LPS was hydrolysed under mild acid conditions and a fraction that contained one O-chain repeating unit linked to a Kdo residue was isolated and characterized by FTICR-ESIMS and NMR spectroscopy. Moreover, after an alkaline reductive hydrolysis, a disaccharide alpha-Kdo-(2 -> 6)-GlcNol was isolated and characterized. The data obtained proved the presence of an alpha-Kdo in the outer core and allowed the identification of the O-antigen biological repeating unit as well as its linkage with the core oligosaccharide. (C) 2010 Elsevier Ltd. All rights reserved.
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