Analysis of lipopolysaccharide (lipid A) fatty acids

Wollenweber, H. W.; Rietschel, Ernest T.

doi:10.1016/0167-7012(90)90056-c

Cited by 161 publications

(111 citation statements)

References 70 publications

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“…Total fatty acids were identified and quantified by transesterification, using acid-catalyzed methanolysis (41 (43). Amino sugars were N-acetylated after methanolysis and prior to trimethylsilylation (43).…”

Section: Materuils and Methodsmentioning

confidence: 99%

Chemical characterization of pH-dependent structural epitopes of lipopolysaccharides from Rhizobium leguminosarum biovar phaseoli

Bhat

Carlson

1992

J Bacteriol

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Lipopolysaccharide (LPS) was isolated from free-living Rhizobium leguminosarum bv. phaseoli CE3 cells grown at pH 4.8 (antigenically similar to bacteroid LPS) and compared with that from cells grown at pH 7.2 (free-living bacteria). Composition analysis revealed that pH 7.2 LPS differs from pH 4.8 LPS in that 2,3,4-tri-O-methylfucose is replaced by 2,3-di-0-methylfucose. The amount of 2-O-methylrhamnose is greater in the pH 4.8 LPS than in the pH 7.2 LPS. Analysis of the structural components of LPS (0-chain polysaccharide, core oligosaccharides, and the lipid A) revealed that all the composition differences in the various LPSs occur in the 0-chain polysaccharide. These structural variations between pH 4.8 and pH 7.2 LPSs provide a chemical basis for the observed lack of cross-reactivity with pH 4.8 LPS of two monoclonal antibodies, JIM28 and JIM29, raised against free-living bacteria grown at pH 7.2. An LPS preparation isolated from bacteroids contained both 2,3,4-tri-O-and 2,3-di-O-methylfucose residues. This result is consistent with the finding that the two monoclonal antibodies react weakly with bacteroid LPS. It is concluded that methylation changes occur on the LPS 0-chain of R. leguminosarum bv. phaseoli when the bacteria are grown at low pH and during nodule development.Rhizobia are gram-negative soil bacteria that form nitrogen-fixing symbiotic associations with leguminous plants. In terms of host selection, the bacteria exhibit a great deal of specificity. The surface carbohydrates of rhizobia, which include the lipopolysaccharide (LPS), extracellular polysaccharide, and capsular polysaccharide, have all been hypothesized to play a role in symbiosis (9,11,18,20,22,23). The LPSs of Rhizobium species resemble their enterobacterial counterparts in having structurally distinct regions; the 0-chain polysaccharide, the core oligosaccharide, and the hydrophobic lipid A (10, 13-17); however, there are many differences in the structural details of Rhizobium LPSs compared with enterobacterial LPSs (1-3, 12, 25-27).Earlier reports on the role of LPSs in symbiosis were directed toward the possibility that LPSs were involved in the specific attachment of the symbiont to the host plant (8,9,30). Recent studies indicate involvement of LPSs at a later stage in the nodule development. Mutants which do not have the 0-chain region either fail to form normal infection threads or are not released properly from the infection thread into the host root cells (7,13,19,33,34,37).Using monoclonal antibodies to Rhizobium leguminosarum bv. viciae bacteroids, it has been shown that there are changes in LPS epitopes which occur during differentiation of bacteria into bacteroids (6,35,39,42). Some of these changes can be produced ex planta, by growing the bacteria at low pH or low 02 tension, conditions which mimic, in part, the microenvironment of the root nodule (28). Similar results have been obtained for another strain of R. leguminosarum bv. viciae and for R leguminosarum bv. phaseoli (21, 38). These epitope changes occur only ...

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Section: Materuils and Methodsmentioning

confidence: 99%

Chemical characterization of pH-dependent structural epitopes of lipopolysaccharides from Rhizobium leguminosarum biovar phaseoli

Bhat

Carlson

1992

J Bacteriol

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“…Among these FAs detected, 18:2ω9c can be used as fungal signature FA,and 12:0 2OH,12:0 3OH,i15:0,a15:0,15:0,14:0 3OH,i16:0,16:1ω9,i17:0,cy17:0,17:0,18:1ω9c,18:1ω9t, and cy19:0 are indicative of bacteria according to previous references (Tunlid et al 1989;Frostegård et al 1993). Regarding diagnostic FAs of bacteria, branched FAs i15:0, a15:0, i16:0, and i17:0 are employed as general indicators of Gram-positive (G + ) bacteria (Rajendran et al 1997), while 12:0 2OH, 12:0 3OH, 14:0 3OH, 16:1ω9, cy17:0, 18:1ω9c, 18:1ω9t, and cy19:0 are primarily associated with Gram-negative (G − ) bacteria (Wollenweber and Rietschel 1990;Zelles 1999b). In addition, monounsaturated fatty acids (MUFAs) are typically contained in aerobic bacteria, whereas branched FAs are primarily contained in anaerobic bacteria; therefore, the ratio of MUFAs/branched FAs can be indicative of the relative amounts of aerobic to anaerobic bacteria (Rajendran et al 1992;Rajendran et al 1997).…”

Section: Signature Fasmentioning

confidence: 99%

Comparative study of microbial community structure in integrated vertical-flow constructed wetlands for treatment of domestic and nitrified wastewaters

Chang

Liang

et al. 2014

Environ Sci Pollut Res

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Microbial processes play a vital important role in the removal of contaminants in constructed wetland (CW). However, the microbial physiology and community structure can be influenced by environmental conditions. In this study, four pilot-scale integrated vertical-flow constructed wetlands (IVCWs) were employed to treat domestic and nitrified wastewaters. The microbial properties, along with their response to wastewater quality characteristics and seasonal variation, were determined. The results showed higher Shannon-Weiner diversity (H) and evenness (E) index of fatty acids (FAs), and relative abundances of signature FAs in down-flow cells and in the systems fed with domestic wastewater (DW). The relative abundances of fungi and gram-negative and aerobic bacteria were greater in up-flow cells. The dominant anaerobic bacteria found in most cells might be accounted for the prevailing anaerobic environment within the wetland beds, which could mean that the system fed with nitrified wastewater (NW) should perform better in nitrogen removal. The redundancy analysis (RDA) showed that pollutant concentrations, especially organic matter, influence the FA compositions greatly, and the most significant difference of microbial community structures was detected in down-flow cells fed with DW and up-flow ones with NW. The branched FAs, which could be used to represent anaerobic bacteria, were observed in down-flow cells treating DW and had a significant positive correlation with chemical oxygen demand (COD) concentration, probably suggesting the important role of anaerobic bacteria in organic matter degradation in the IVCWs. Seasonal variation, however, did not greatly influence the microbial community structure in the IVCWs.

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“…The absolute configuration of the hydroxy fatty acids was determined by GLC of the phenylethylamide derivatives (10). Analyses for ester-and amide-linked fatty acids and ester-linked acyloxyacyl groups were performed as described (11), as was SDS-PAGE (10 and 18% acrylamide) (12), gels of which were stained with silver nitrate for the detection of LPS (13) or with Coomassie Brilliant Blue for the detection of proteins. Analytical and semipreparative HPAE were performed as described (14), with the modification that in semipreparative HPAE the CarboPac PA1 column was eluted at 3 ml min…”

Section: Structure Of K Pneumoniae Lipopolysaccharidementioning

confidence: 99%

Identification of a Novel Heptoglycan of α1→2-Linkedd-glycero-d-manno-Heptopyranose

Susskind

Brade

et al. 1998

Journal of Biological Chemistry

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After immunization of BALB/c mice, two monoclonal antibodies were obtained that were shown to be specific for the core of LPS from K. pneumoniae ssp. pneumoniae, since they did not react with LPS or whole-cell lysates of a variety of other Gram-negative species. Both monoclonal antibodies could be inhibited by LPS but not by isolated oligosaccharides and are thus considered to recognize a conformational epitope in the core region.

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Analysis of lipopolysaccharide (lipid A) fatty acids

Cited by 161 publications

References 70 publications

Chemical characterization of pH-dependent structural epitopes of lipopolysaccharides from Rhizobium leguminosarum biovar phaseoli

Chemical characterization of pH-dependent structural epitopes of lipopolysaccharides from Rhizobium leguminosarum biovar phaseoli

Comparative study of microbial community structure in integrated vertical-flow constructed wetlands for treatment of domestic and nitrified wastewaters

Identification of a Novel Heptoglycan of α1→2-Linkedd-glycero-d-manno-Heptopyranose

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