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J. Bacteriol. doi:10.1128/JB.00891-08
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Two Kdo-heptose regions identified in Hafnia alvei 32 lipopolysaccharide: the complete core structure and serological screening of different Hafnia O-serotypes

Department of Immunochemistry, L. Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, R. Weigla 12, PL-53-114 Wroclaw, Poland; and Department of Chemistry, Swedish University of Agricultural Sciences, SE-750 07 Uppsala, Sweden

^* To whom correspondence should be addressed. Email: czaja{at}iitd.pan.wroc.pl.

	Abstract

Hafnia alvei, a Gram-negative bacterium, is an opportunistic pathogen associated with mixed hospital infections, bacteremia, septicemia, and respiratory diseases. Various Kdo (3-deoxy-d-manno-oct-2-ulosonic acid)-containing fragments, different from already known structures of core oligosaccharides, were previously found among fractions obtained by mild acid hydrolysis of some H. alvei lipopolysaccharides. However, the position of these segments in LPS structure was not known. Analysis of de-N,O-acylated LPS by NMR spectroscopy and mass spectrometry allowed for determination of the location of Kdo-containing trisaccharide in the structure of H. alvei PCM 32 LPS. It was established that the trisaccharide [l--d-Hepp-(14)-[-d-Galp6OAc-(17)]--Kdop-(2] is an integral part of the outer core oligosaccharide of H. alvei 32 LPS. The ketosidic linkage of very labile nature between trisaccharide's 4,7)--Kdop and 2)-Glcp in the core oligosaccharide was identified. Screening for this Kdo-containing trisaccharide was performed on the group of 37 O-serotypes of H. alvei LPSs using monospecific antibodies recognizing this structure. It was established that this trisaccharide is a characteristic component of the outer core oligosaccharides of H. alvei 2, 32, 600, 1192, 1206, and 1211 LPSs. The weaker cross-reactions with LPSs of strains 974, 1188, 1198, 1204, and 1214 suggest the presence of similar structures in these LPSs as well. Thus we have identified new example of endotoxins among those elucidated so far. This type of core oligosaccharide deviates from the classical scheme by the presence of the structural Kdo-containing motif in the outer core region.