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Journal of Bacteriology, January 2009, p. 533-544, Vol. 191, No. 2
0021-9193/09/$08.00+0     doi:10.1128/JB.00891-08
Copyright © 2009, American Society for Microbiology. 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 ^,

Jolanta Lukasiewicz,^1* Tomasz Niedziela,¹ Wojciech Jachymek,¹ Lennart Kenne,² and Czeslaw Lugowski¹

Department of Immunochemistry, L. Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, R. Weigla 12, PL-53-114 Wroclaw, Poland,¹ Department of Chemistry, Swedish University of Agricultural Sciences, SE-750 07 Uppsala, Sweden²

Received 30 June 2008/ Accepted 5 November 2008

Hafnia alvei, a gram-negative bacterium, is an opportunistic pathogen associated with mixed hospital infections, bacteremia, septicemia, and respiratory diseases. Various 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo)-containing fragments different from known structures of core oligosaccharides were previously found among fractions obtained by mild acid hydrolysis of some H. alvei lipopolysaccharides (LPSs). However, the positions of these segments in the LPS structure were not known. Analysis of de-N,O-acylated LPS by nuclear magnetic resonance spectroscopy and mass spectrometry allowed the determination of the location of a 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 very labile ketosidic linkage between 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 the 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 examples 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.

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* Corresponding author. Mailing address: L. Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, R. Weigla 12, PL-53-114 Wroclaw, Poland. Phone: 48 71 370 99 27. Fax: 48 71 337 13 82. E-mail: czaja{at}iitd.pan.wroc.pl

Published ahead of print on 14 November 2008.

Supplemental material for this article may be found at http://jb.asm.org/.

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Journal of Bacteriology, January 2009, p. 533-544, Vol. 191, No. 2
0021-9193/09/$08.00+0     doi:10.1128/JB.00891-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.