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J. Biol. Chem., Vol. 278, Issue 30, 27502-27512, July 25, 2003

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The Obligate Predatory Bdellovibrio bacteriovorus Possesses a Neutral Lipid A Containing -D-Mannoses That Replace Phosphate Residues

SIMILARITIES AND DIFFERENCES BETWEEN THE LIPID As AND THE LIPOPOLYSACCHARIDES OF THE WILD TYPE STRAIN B. BACTERIOVORUS HD100 AND ITS HOST-INDEPENDENT DERIVATIVE HI100^*

Dominik Schwudke  , Michael Linscheid  ¶, Eckhard Strauch , Bernd Appel , Ulrich Zähringer ||, Hermann Moll ||, Mareike Müller ||, Lothar Brecker ||, Sabine Gronow || and Buko Lindner ||

From the Department of Chemistry, Humboldt Universität zu Berlin, D-12489 Berlin, Germany, the Project Group Biological Safety, Robert Koch Institute Berlin, D-13353 Berlin, Germany, and the ^||Research Center Borstel, Center for Medicine and Biosciences, D-23845 Borstel, Germany

Bdellovibrio bacteriovorus are predatory bacteria that penetrate Gram-negative bacteria and grow intraperiplasmically at the expense of the prey. It was suggested that B. bacteriovorus partially degrade and reutilize lipopolysaccharide (LPS) of the host, thus synthesizing an outer membrane containing structural elements of the prey. According to this hypothesis a host-independent mutant should possess a chemically different LPS. Therefore, the lipopolysaccharides of B. bacteriovorus HD100 and its host-independent derivative B. bacteriovorus HI100 were isolated and characterized by SDS-polyacrylamide gel electrophoresis, immunoblotting, and mass spectrometry. LPS of both strains were identified as smooth-form LPS with different repeating units. The lipid As were isolated after mild acid hydrolysis and their structures were determined by chemical analysis, by mass spectrometric methods, and by NMR spectroscopy. Both lipid As were characterized by an unusual chemical structure, consisting of a -(16)-linked 2,3-diamino-2,3-dideoxy-D-glucopyranose disaccharide carrying six fatty acids that were all hydroxylated. Instead of phosphate groups substituting position O-1 of the reducing and O-4' of the nonreducing end -D-mannopyranose residues were found in these lipid As. Thus, they represent the first lipid As completely missing negatively charged groups. A reduced endotoxic activity as determined by cytokine induction from human macrophages was shown for this novel structure. Only minor differences with respect to fatty acids were detected between the lipid As of the host-dependent wild type strain HD100 and for its host-independent derivative HI100. From the results of the detailed analysis it can be concluded that the wild type strain HD100 synthesizes an innate LPS.

Received for publication, March 24, 2003 , and in revised form, May 7, 2003.

^* The work was supported by Deutsche Forschungsgemeinschaft Grants LI 448/1-1 (to B. L.), SFB 470-A1 (to S. G.), SFB 470-B4 (to U. Z.), and LI 309/22-1 (to M. L.) and the Fonds der Chemischen Industrie (FCI). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ To whom correspondence should be addressed: Professor of Analytical Chemistry, Humboldt Universität zu Berlin, Dept. of Chemistry, Brook-Taylor-Str. 2, 12489 Berlin, Germany. Tel.: 49-0-30-2093-7575; Fax: 49-0-30-2093-6985; E-mail: michael.linscheid{at}chemie.hu-berlin.de.

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