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Chemical composition of lipopolysaccharides from Legionella bozemanii and Legionella longbeachae

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Abstract

Lipopolysaccharides (LPS) from Legionella bozemanii serogroup 1 and Legionella longbeachae serogroup 1 were subjected to chemical analyses. The lipid A part of both LPSs contained 2,3-dideoxy-2,3-diamino-d-glucose as major constituents and d-glucosamine and glycerol as minor constituents of the sugar backbone structure. Both LPSs exhibited a very complex fatty acid composition. Twenty amide-linked 3-hydroxy fatty acids were detected in LPS of L. longbeachae, whereas seventeen were encountered in LPS of L. bozemanii. Both LPSs contained nine ester-linked nonhydroxy fatty acids and the unique long-chain fatty acids 27-oxo-octacosanoic acid, 29-oxotriacontanoic acid, heptacosane-1,27-dioic acid and nonacosane-1,29-dioic acid. SDS-PAGE showed that L. bozemanii produced smooth-form LPS, whereas L. longbeachae LPS was mainly of the R-type. Composition analyses were in accordance with these electrophoretic patterns. d-Quinovosamine and l-fucosamine constituted 80 mol% of the polysaccharide part of L. bozemanii LPS. Other sugars identified were d-glucosamine, d-mannose, d-glucose, l-rhamnose, d-glycero-d-manno-heptose, l-glycero-d-mannoheptose, 2-keto-3-deoxy-octonic acid and glycerol. The polysaccharide chain from LPS of L. longbeachae appeared to be shorter, but composed of the same sugars except l-fucosamine. Both LPSs contained glycerol phosphate and glucosamine phosphate and L. longbeachae LPS contained in addition glucose phosphate.

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Abbreviations

EI :

Electron impact

GlcN3N :

2,3-Diamino-2,3-dideoxy-d-glucose

HPAEC :

High pH anion-exchange chromatography

Kdo :

2-Keto-3-deoxy-octonic acid

LPS :

Lipopolysaccharide

PCP :

Phenol/chloroform/petroleum ether solvent

PED :

Pulsed electrochemical detection

PS :

Polysaccharide

TFA :

Trifluoroacetyl

TMS :

Trimethylsilyl

References

  • Arata S, Shiga T, Mizutani T, Mashimo J, Kasai N, Szabo L (1992) Structure of a new type of lipid A from Pseudomonas vesicularis (abstract). 2nd Conference of the International Endotoxin Society, Aug. 17–20, Vienna, Austria

  • Bhat UR, Carlson RW, Busch M, Mayer H (1991) Distribution and phylogenetic significance of 27-hydroxyoctacosanoic acid in lipopolysaccharides from bacteria belonging to the alpha-2 subgroup of Proteobacteria. Int J Syst Bacteriol 41:213–217

    Google Scholar 

  • Bitter T, Muir HM (1962) A modified uronic acid carbazole reaction. Anal Biochem 4:330–334

    Google Scholar 

  • Brade H, Galanos C, Lüderitz O (1983) Differential determination of the 3-deoxy-d-mannooctulosonic acid residues in lipopolysaccharides of Salmonella minnesota rough mutants. Eur J Biochem 131:195–200

    Google Scholar 

  • Brenner DJ (1987) Classification of the Legionellae. Semin Respir Infect 2:190–205

    Google Scholar 

  • Bryn K, Jantzen E (1986) Quantification of 2-keto-3-deoxy-octo-nate in (lipo)polysaccharides by methanolytic release, trifluoroacetylation and capillary gas chromatography. J Chromatogr 370:103–112

    Google Scholar 

  • Collins MD, Gilbart J (1983) New members of the coenzyme Q series from the Legionellaceae. FEMS Microbiol Lett 16:251–255

    Google Scholar 

  • Dennis PJ, Brenner DJ, Thacker WL, Wait R, Vesey G, Steigerwalt AG, Benson RF (1993) Five new Legionella species isolated from water. Int J Sys Bacteriol 43:329–337

    Google Scholar 

  • Fallon RJ (1990) Legionella. In: Parker MT, Duerden BI (eds) Systematic bacteriology, vol 2. Edward Arnold, London, pp 275–287

    Google Scholar 

  • Fang GD, Yu VL, Vickers RM (1989) Disease due to the Legionellaceae (other than Legionella pneumophila). Medicine (Baltimore). 68:116–132

    Google Scholar 

  • Fox A, Rogers JC, Fox KF, Schnitzer G, Morgan SL, Brown A, Aono R (1990) Chemotaxonomic differentiation of Legionellae by detection and characterization of aminodideoxyhexoses and other unique sugars using gas chromatography-mass spectrometry. J Clin Microbiol 28:546–552

    Google Scholar 

  • Fry NK, Warwick S, Saunders NA, Embley TM (1991) The use of 16S ribosomal RNA analyses to investigate the phylogeny of the family Legionellaceae. J Gen Microbiol 137:1215–1222

    Google Scholar 

  • Galanos C, Lüderitz O, Westphal O (1979) Preparation and properties of a standardized lipopolysaccharide from Salmonella abortus equi (Novo-Pyrexal). Zentralbl Bakteriol Hyg I [A] 243:226–244

    Google Scholar 

  • Gerwig GJ, Kamerling JP, Vliegenthart JFG (1978) Determination of the d and l configuration of neutral monosaccharides by high-resolution capillary GLC. Carbohydr Res 62:349–357

    Google Scholar 

  • Hollingsworth RI, Lill-Elghanian DA (1989) Isolation and characterization of the unusual lipopolysaccharide component, 2-amino-2-deoxy-2-N-(27-hydroxyoctacosanoyl)-3-O-(3-hydroxytetradecanoyl)-gluco-hexuronic acid, and its de-O-acylation product from the free lipid A of Rhizobium trifolii ANU843. J Biol Chem 264:14039–14042

    Google Scholar 

  • Jantzen E, Sonesson A, Tangen T, Eng J (1993) Hydroxy-fatty acid profiles of Legionella species: diagnostic usefulness assessed by principal component analysis. J Clin Microbiol 31: 1413–1419

    Google Scholar 

  • Johnson KG, Perry MB (1976) Improved techniques for the preparation of bacterial lipopolysaccharides. Can J Microbiol 22: 29–34

    Google Scholar 

  • Kawahara K, Brade H, Rietschel ET, Zähringer U (1987) Studies on the chemical structure of the core-lipid A region of the lipopolysaccharide of Acinetobacter calcoaceticus NCTC 10305. Detection of a new 2-octulosonic acid interlinking the core oligosaccharide and lipid A component. Eur J Biochem 163:489–495

    Google Scholar 

  • Kickhöfen B, Warth R (1968) Eine Trennkammer für die Hochspannungselektrophorese nach den Michl' schen Prinzip. J Chromatogr 33:558–560

    Google Scholar 

  • Lambert MA, Moss CW (1989) Cellular fatty acid compositions and isoprenoid quinone contents of 23 Legionella species. J Clin Microbiol 27:465–473

    Google Scholar 

  • Lesse AJ, Campagnari AA, Bittner WE, Apicella MA (1990) Increased resolution of lipopolysaccharides and lipooligosaccharides utilizing tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis. J Immunol Methods 126:109–117

    Google Scholar 

  • Lindberg B (1990) Components of bacterial polysaccharides. In: Tipson RS, Horton D (eds) Advances in carbohydrate chemistry and biochemistry, vol 48. Academic Press, San Diego, pp 279–318

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275

    Google Scholar 

  • Lowry OH, Roberts NR, Leiner KY, Wu ML, Farr AL (1954) The quantitative histochemistry of brain. I. Chemical methods. J Biol Chem 207:1–17

    Google Scholar 

  • L'vov V, Gur'yanova SV, Rodionov AV, Gorshkova RP (1992) Structure of the repeating unit of the O-specific polysaccharide of the lipopolysaccharide of Yersinia kristensenii strain 490 (O:12,25). Carbohydr Res 228:415–422

    Google Scholar 

  • Mayberry WR (1984) Monohydroxy and dihydroxy fatty acid composition of Legionella species. Int J Sys Bacteriol 34:321–326

    Google Scholar 

  • Mayer H, Krauss JH, Yokota A, Weckesser J (1990) Natural variants of lipid A. In: Friedman H, Klein TW, Nakano M, Nowotny A (eds) Endotoxin. Plenum, New York, pp 45–70

    Google Scholar 

  • Moll H, Sonesson A, Jantzen E, Marre R, Zähringer U (1992) Identification of 27-oxo-octacosanoic acid and heptacosane-1,27-dioic acid in Legionella pneumophila. FEMS Microbiol Lett 97:1–6

    Google Scholar 

  • Moran AP, Zähringer U, Seydel U, Scholz D, Stütz P, Rietschel ET (1991) Structural analysis of the lipid A component of Campylobacter jejuni CCUG 10936 (serotype O:2) lipopolysaccharide. Description of a lipid A containing a hydrid backbone of 2-amino-2-deoxy-d-glucose and 2,3-diamino-2,3-dideoxy-d-glucose. Eur J Biochem 198:459–469

    Google Scholar 

  • Reisner AH, Nemes P, Bucholtz C (1975) The use of Coomassie brilliant blue G250 perchloric acid solution for staining in electrophoresis and isoelectric focusing on polyacrylamide gels. Anal Biochem 64:509–516

    Google Scholar 

  • Roppel J, Mayer H, Weckesser J (1975) Identification of a 2,3-diamino-2,3-dideoxyhexose in the lipid A component of lipopolysaccharides of Rhodopseudomonas viridis and Rhodopseudomonas palustris. Carbohydr Res 40:31–40

    Google Scholar 

  • Schägger H, Jagow G von (1987) Tricine-sodium dodecyl sulfatepolyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100kDa. Anal Biochem 166: 368–379

    Google Scholar 

  • Sonesson A, Jantzen E (1992) The branched-chain octose yersiniose A is a lipopolysaccharide constituent of Legionella micdadei and Legionella maceachernii. J Microbiol Methods 15: 241–248

    Google Scholar 

  • Sonesson A, Larsson L, Fox A, Westerdahl G, Odham G (1988) Determination of environmental levels of peptidoglycan and lipopolysaccharide using gas chromatography with negativeion chemical-ionization mass spectrometry utilizing bacterial amino acids and hydroxy fatty acids as biomarkers. J Chromatogr 431:1–15

    Google Scholar 

  • Sonesson A, Bryn K, Jantzen E, Larsson L (1989a) Gas chromatographic determination of (phosphorylated) 2-keto-3-deoxyoctonic acid, heptoses and glucosamine in bacterial lipopolysaccharides after treatment with hydrofluoric acid, methanolysis and trifluoroacetylation. J Chromatogr 487:1–7

    Google Scholar 

  • Sonesson A, Jantzen E, Bryn K, Larsson L, Eng J (1989b) Chemical composition of a lipopolysaccharide from Legionella pneumophila. Arch Microbiol 153:72–78

    Google Scholar 

  • Sonesson A, Jantzen E, Bryn K, Tangen T, Eng J, Zähringer U (1994) Composition of 2,3-dihydroxy fatty acid-containing lipopolysaccharides from Legionella israelensis, Legionella maceachernii and Legionella micdadei. Microbiology 140:1261–1271

    Google Scholar 

  • Tsai C-M, Frasch CE (1982) A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem 119: 115–119

    Google Scholar 

  • Waravdekar VS, Saslaw LD (1959) A sensitive colorimetric method for the estimation of 2-deoxy sugars with the use of the malonaldehyde-thiobarbituric acid reaction. J Biol Chem 234: 1945–1950

    Google Scholar 

  • Westphal O, Lüderitz O, Bister F (1952) Über die extraktion von Bakterien mit Phenol/Wasser. Z Naturforsch [B] 7:148–155

    Google Scholar 

  • Wilkinson SG, Galbraith L, Lightfoot GA (1973) Cell walls, lipids, and lipopolysaccharides of Pseudomonas species. Eur J Biochem 33:158–174

    Google Scholar 

  • Zamze SE, Ferguson MAJ, Moxon ER, Dwek RA, Rademacher TW (1987) Identification of phosphorylated 3-deoxy-manno-octulosonic acid as a component of Haemophilus influenzae lipopolysaccharide. Biochem J 245:583–587

    Google Scholar 

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Author notes

  1. Anders Sonesson

    Present address: Astra Draco AB, P.O. Box 34, S-22100, Lund, Sweden

Authors and Affiliations

  1. National Institute of Public Health, Geitmyrsveien 75, N-0462, Oslo, Norway

    Anders Sonesson, Erik Jantzen & Torill Tangen

  2. Forschungsinstitut Borstel, Institut für Experimentelle Biologie und Medizin, Parkallee 22, D-23845, Borstel, Germany

    Ulrich Zähringer

Authors
  1. Anders Sonesson
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  2. Erik Jantzen
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  3. Torill Tangen
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  4. Ulrich Zähringer
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Sonesson, A., Jantzen, E., Tangen, T. et al. Chemical composition of lipopolysaccharides from Legionella bozemanii and Legionella longbeachae . Arch. Microbiol. 162, 215–221 (1994). https://doi.org/10.1007/BF00301841

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  • DOI: https://doi.org/10.1007/BF00301841

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