Occurrence of lipid A variants with 27-hydroxyoctacosanoic acid in lipopolysaccharides from members of the family Rhizobiaceae (original) (raw)
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Journal of Biological Chemistry, 1994
The structure of lipid A from the lipopolysaccharide of Rhizobium leguminosarum bv.phaseoli (wild type strain CE3) was investigated by alkylation analysis, nuclear magnetic resonance spectroscopy, and electrospray and fast atom bombardment mass spectrometry of the de-0acylated lipid A. The lipid A carbohydrate backbone was shown to be a trisaccharide containing galacturonic acid, glucosamine, and the unique sugar 2-amino-2-deoxygluconic acid, previously unreported in lipopolysaccharides. Nuclear magnetic resonance spectroscopy and ethylation analyses revealed that the galacturonic acid is a-1,Clinked to the glucosamine, while the amino aldonic acid residue, which may exist as the l,&lactone, is attached as an aglycone to the glucosamine and, thus, occupies the reducing end of the molecule. The resulting backbone is hydrophilic and analogous to the commonly observed bisphosphorylated glucosamine disaccharide from enteric bacterial lipopolysaccharides in that both the nonreducing and reducing ends carry negatively charged substituents. The fatty acids of the R. leguminosarum lipid A are attached both as 0-and N-acyl substituents to glucosamine and 2-aminogluconate. All fatty acids are hydroxylated consisting of 3-hydroxymyristate (3-0H-Cl4.,), 3-hydroxypentadecanoate (3-OH-Cis.,), 3-hydroxypalmitate (3-OH-C16,), 3-hydroxystearate (3-OH-C18,), and 27-hydroxyoctacosanoate (27-OH-C,,,,) in the approximate mole ratio 3:0.2:1:0.61. Unlike lipid As from enteric bacteria, the R. leguminosarum lipid A lacks 3-acyloxyacyl substituents; however, the long chain 27-hydroxy fatty acid carries ester-linked P-hydroxybutyrate at the 27-hydroxy position. Fast atom bombardment mass spectrometry of the de-0-acylated lipid A demonstrated the presence of 2 molecular species that differ by 28 mass units due to fatty acid heterogeneity at the two amide linkages. One species carries amide-linked 3-OH
Chemical characterization of two lipopolysaccharide species isolated from Rhizobium loti NZP2213
Archives of Microbiology, 1995
Phenol-water extraction of Rhizobium loti NZP2213 cells allowed a simultaneous isolation of two structurally different lipopolysaccharides, from the aqueous (LPS-W) and phenol (LPS-P) phase that differed in their sodium doexycholate-PAGE pattern and composition. LPS-W showed a profile indicating an R-type LPS; LPS-P had a cluster of poorly resolved bands in the high-molecular-weight region. LPS-P contained large amounts of 6-deoxy-l-talose (6dTal), and a small amount of 2-O-methyl-6-deoxy-talose (molar ratio ≈30:1), both of which were completely absent in LPS-W. Methylation analysis gave only one major product, 2,4-di-O-methyl-6dTal, indicating that the O-chain is composed of a homopolymer of 1,3-linked 6dTal, having the methylated 6dTal (2-O-me-6dTal) probably localized at the non-reducing end of the O-chain. This homopolymeric O-chain was additionally O-acetylated, as evidenced by GC-MS and by 13C NMR analysis. The lipid A moieties of both LPS-W and LPS-P showed almost identical composition, with six, different 3-OH fatty acids and with two, so far not described, long-chain 4-oxo-fatty acids, all being amide-linked, and with 27-OH-28:0 as the main ester-linked fatty acid. Lipid A was of the lipid ADAG-type, i.e., having a (phosphorylated) 2,3-diamino-2,3-dideoxy-d-glucose-containing lipid A backbone. Lipid ADAG is widespread among species of the α-2 group of Proteobacteria, but has so far not been encountered in any other rhizobial or agrobacterial species.
Structure of lipid A from a stem-nodulating bacterium Azorhizobium caulinodans
Carbohydrate Research, 2012
The structure of the lipid A of the lipopolysaccharide (LPS) from Azorhizobium caulinodans, a symbiont of the tropical legume Sesbania rostrata, was investigated by chemical compositional analyses, mass spectrometry, as well as 1D and 2D NMR spectroscopy techniques. The lipid A backbone was composed of a b-(1?6)-linked 2,3-diamino-2,3-dideoxy-D-glucopyranose (GlcpN3N) disaccharide and a-D-glucuronic acid (GlcpA). Nuclear magnetic resonance spectroscopy revealed that the GlcpA was connected to the reducing end of the diaminosugar disaccharide via an a-(1?1) glycosidic bond. The lipid A was deprived of phosphate residues. ESI-MS analysis showed that the lipid A preparation was a mixture of molecules due to the occurrence of different acylation patterns. The GlcpN3N disaccharide backbone was N-acylated at the C-2, C-3, C-2 0 and C-3 0 positions with 3-OH-18:0, 3-OH-14:0, 3-OH-20:1 and 3-OH-14:0 fatty acids, respectively. Nonpolar fatty acids as well as 3-OH-18:0 were found to be ester-linked. They were attached to hydroxyl groups of primary 3-OH fatty acids giving three acyloxyacyl moieties. Thus, the complete lipid A from A. caulinodans comprised seven acyl residues. Part of the lipid A molecules was esterified by 3-methoxybutyric acid. Azorhizobium caulinodans did not incorporate x-1 hydroxylated very long chain fatty acids (e.g., 27-OH-28:0) into the lipid A, which makes this variant of endotoxin unusual among rhizobial lipids A.
The Journal of biological chemistry, 2014
The chemical structures of the unusual hopanoid-containing lipid A samples of the lipopolysaccharides (LPS) from three strains of Bradyrhizobium (slow-growing rhizobia) have been established. They differed considerably from other Gram-negative bacteria in regards to the backbone structure, the number of ester-linked long chain hydroxylated fatty acids, as well as the presence of a tertiary residue that consisted of at least one molecule of carboxyl-bacteriohopanediol or its 2-methyl derivative. The structural details of this type of lipid A were established using one- and two-dimensional NMR spectroscopy, chemical composition analyses, and mass spectrometry techniques (electrospray ionization Fourier-transform ion cyclotron resonance mass spectrometry and MALDI-TOF-MS). In these lipid A samples the glucosamine disaccharide characteristic for enterobacterial lipid A was replaced by a 2,3-diamino-2,3-dideoxy-d-glucopyranosyl-(GlcpN3N) disaccharide, deprived of phosphate residues, and ...
Journal of Bacteriology, 2003
The structure of the lipid A from Rhizobium etli and Rhizobium leguminosarum lipopolysaccharides (LPSs) lacks phosphate and contains a galacturonosyl residue at its 4′ position, an acylated 2-aminogluconate in place of the proximal glucosamine, and a very long chain ω-1 hydroxy fatty acid, 27-hydroxyoctacosanoic acid (27OHC28:0). The 27OHC28:0 moiety is common in lipid A's among members of the Rhizobiaceae and also among a number of the facultative intracellular pathogens that form chronic infections, e.g., Brucella abortus , Bartonella henselae , and Legionella pneumophila . In this paper, a mutant of R. leguminosarum was created by placing a kanamycin resistance cassette within acpXL , the gene which encodes the acyl carrier protein for 27OHC28:0. The result was an LPS containing a tetraacylated lipid A lacking 27OHC28:0. A small amount of the mutant lipid A may contain an added palmitic acid residue. The mutant is sensitive to changes in osmolarity and an increase in acidity,...
Systematic and Applied Microbiology, 2000
Lipopolysaccharides (LPSs) isolated from seven strains of Mesorhizobium were studied for the presence of fatty acids with particular attention for 27-oxooctacosanoic acid and 4-oxo fatty acids. The LPSs from all analysed strains contained various amounts of 27-oxo-28:0 and all of them, with the exception of Mesorhizobium tianshanense, contained also 4-oxo fatty acids (4-oxo-20:0, 4-oxo-i-21:0, 4-oxo-22:0). The group of amide-linked fatty acids consisted of a wide range of 3-hydroxylated and 4-oxo fatty acids whereas all the nonpolar as well as the (m-l) hydroxylated long-chain acids and the 27-oxo-28:0 fatty acids were ester-linked. The characteristic spectrum of 3-hydroxy fatty acids and presence of 27-0H-28:0 as well as 27-oxo-28:0 acid in LPSs of Mesorhizobium showed that these strains were closely related. Therefore the lipid A fatty acid pattern could be a useful chemotaxonomic marker which helps to isolate the Mesorhizobium group from rhizobium bacteria during the classification process.
Lipid-bound sugars in Rhizobium meliloti
1980
Incubation of an enzyme preparation of Rhizobium meliloti with labeled uridine diphosphate glucose led to the formation of radioactive substances soluble in organic solvents. These substances are probably polyprenyl diphosphate saccharides. They behaved like these on treatment with ammonia or with hot phenol and were decomposed by heating for 10 min at pH 2 yielding a mono-and a disaccharide. The monosaccharide was identified as galactose by paper chromatography. The disaccharide gave glucose and galactose by acid hydrolysis. Following reduction with borohydride it yielded glucose and galactitol. After treatment with periodate followed by paper chromatography only galactose was detectable. The disaccharide was hydrolyzed by p-but not by cY-glucosidase. Therefore the disaccharide is glucosyl pl-3-galactose.
Systematic and Applied Microbiology, 1996
Structural studies of two lipopolysaccharides (LPS) isolated from Rhizobium leguminosarum bv. trifolii strain 24 (giving effective symbiosis with clover) and the exo-mutant AR20 showing non-effective nodulation, were performed using conventional techniques (chemical analysis, gas chromatography (GC), methylation analysis, combined gas chromatography/mass spectrometry (GC-MS)) and NMR. The two LPS preparations contained: L-rhamnose, 6-deoxy-L-talose (dTal), D-mannose, D-galactose, D-glucose, 3-deoxy-2-heptulosaric acid (DHA) and 2-keto-3-deoxyoctonate (KDO) as major sugar components, as well as a 2,3-di-O-methylhexose, a 2,6-dideoxy-2-amino-hexose, D-glucosamine and glucosaminic acid as minor constituents. D-galacturonic acid (GaiA), however, was found only in the preparation from the wild-type strain 24. All neutral hexoses and 6-deoxyhexoses have been identified as a-linked anomers. The detailed elucidation of two trisaccharide, derived from the exo-mutant AR20, namely a-L-dTalp(1~2)a-L-Rhap(1~5)DHA and aD -Galp(l~5)a-D-Manp(1~6)KDO by methylation analysis and NMR techniques will be presented. Furthermore, the overall structure of the LPS of the wild-type strain 24 will be proposed.