The lipopolysaccharide of the green sulfur bacterium Chlorobium vibrioforme f. thiosulfatophilum

The cell wall lipopolysaccharide of the green sulfur bacterium Chlorobium vibrioforme f. thiosulfatophilum was obtained by the phenol-chloroform-petroleum ether and the hot phenol-water methods, respectively. It contained mannose, glucose, galacturonic acid, glucosamine, glycine, and small amounts of rhamnose, galactose and glucuronic acid. In addition to d-glycero-d-mannoheptose, the corespecific constituents 2-keto-3-deoxyoctonate and l-glycero-d-mannoheptose were found. Polyacrylamide gel-electrophoresis in the presence of sodium deoxycholate gave no indication for the presence of O-specific repeating units. Degradation of the lipopolysaccharide required 10% acetic acid (100° C, 2 h). The lipid A moiety contained the total of glucosamine of the lipopolysaccharide as well as small amounts of 2,3-diamino-2,3-dideoxy-glucose. It was phosphate-free. The fatty acid spectrum comprised 3-OH-14:0, 3-OH-16:0, and iso-3-OH-18:0 besides little 12:0, 14:0 and 16:0. Hydroxylaminolysis and sodium methylate treatment revealed all of the three hydroxy fatty acids to be amidebound.Abbreviations DOC sodium deoxycholate - PAGE polyacrylamide gel-electrophoresis     

Chemical and biological studies on the lipopolysaccharide (O-antigen) of Anacystis nidulans

The O-antigen (lipopolysaccharide) of Anacystis nidulans, strain KM, has been isolated from whole cells and from cell wall preparations by phenolwater extraction. The polysaccharide moiety consists of a D-mannose polymer accompanied by smaller amounts of 3- and 4-O-methyl-D-mannoses, D-galactose, D-glucose, L-fucose, D-glucosamine, mannosamine and 2-keto-3-deoxyoctonate. Aldoheptoses are lacking. The degraded polysaccharide is split from lipid A by acid hydrolysis (10% acetic acid, 100°C, 3 h) whereby 2-keto-3-deoxyoctonate is released in small amounts. Degraded polysaccharide forms only one major fraction by Sephadex G-50 gel-filtration. This fraction includes all the sugars mentioned above except L-fucose, which is released during the acetic acid degradation. Periodate studies and methylation analysis revealed that the poly-mannose chain consists of about 75% 13 linked and of 25% 14 linked D-mannose units.Lipid A of A. nidulans is phosphate-free. The main fatty acid, -hydroxypalmitic acid, is exclusively amide-bound, presumably to the amino group of D-glucosamine. Other fatty acids, found as minor constituents, are -hydroxymyristic, palmitic and stearic acids. Lipopolysaccharide of A. nidulans KM exhibits high anticomplementary activity in guineapig serum. It is about 800 times less toxic for adrenalectomized mice than endotoxin from Salmonella typhimurium.The isolated lipopolysaccharide reacts with rabbit antisera against living or heat-killed cells of A. nidulans in passive hemagglutination, when untreated or heated, but not when alkali-treated lipopolysaccharide is used for red blood cell sensibilization. It is concluded that lipopolysaccharide of A. nidulans KM is exposed on the surface of the cell.     

The lipopolysaccharide of the phototrophic bacterium Ectothiorhodospira vacuolata

The lipopolysaccharide of Ectothiorhodospira vacuolata was obtained by the phenol-water procedure. It contained a 3-O-methyl-hexose, glucose, galacturonic and glucuronic acids. The finding of d-glycero-d-mannoheptose and 2-keto-3-deoxyoctonate (tentatively identified) suggested a core-structure. The lipid fraction of the lipopolysaccharide contained phosphate and both, 2,3-diamino-2,3-dideoxy-d-glucose and d-glucosamine. The major fatty acids were amine-bound 3-OH-10:0 and 3-OH-12:0 and esterbound 14:0 and 16:0 Sodium deoxycholate gel-electrophoresis, showing a single band only, indicated R-type character of the lipopolysaccharide of Ectothiorhodospira vacuolata.Abbreviations DOC sodium deoxycholate - GC/MS combined gasliquid chromatography - PAGE polyacrylamide gel-electrophoresis     

Extraction and biochemical analyses ofHelicobacter pylori lipopolysaccharides

Lipopolysaccharides were isolated from dehydratedHelicobacter pylori cells by the phenol-chloroform-petroleum ether and hot phenol/water extraction techniques. Biochemical characterization of the crude extracts indicated the following: The primary fatty acids and approximate molar ratios were 3-hydroxyoctadecanoic (2), 3-hydroxyhexadecanoic (1), and octadecanoic (1) acids. Lesser amounts of tetradecanoic, hexadecanoic, and octadecenoic acids were noted. 3-Hydroxytetradecanoic acid was not deteted in either extract. Neutral sugar analyses detected glucose, galactose, two heptose isomers, and an unidentified deoxy-hexose. Glucosamine and glucosamine phosphate were the only amino sugars found in significant quantities. Analyses of other components included ethanolamine, phosphate, and protein. 3-Deoxy-d-manno-octulosonic acid (KDO) was detected, but in lower concentrations than would be expected in comparable enterobacterial lipopolysaccharides.     

Chemical analysis of Azospirillum lipopolysaccharides

Lipopolysaccharides (LPS) were extracted by hot phenol-water from five strains each of Azospirillum lipoferum and Azospirillum brasilense. Rhamnose, glucose, glucosamine and 3-deoxy-d-mannooctulosonic acid were comon sugar constituents of all LPS preparations. 2-O-Mefucose, 3-O-Me-fucose, 3-O-Me-rhamnose and 2-O-Megalactose were found in LPSs of some A. brasilense strains. Fatty acid spectra from all LPSs studied were almost identical with predominance of 3-hydroxymyristic and 3-hydroxypalmitic acids. 3-Hydroxypalmitic acid was the only amide-linked fatty acid. Lipopolysaccharides isolated from A. brasilense showed higher heterogeneity in sugar composition than those from A. lipoferum.Abbreviations glc gas liquid chromatography - ms mass spectrometry - LPS lipopolysaccharide - dOclA 3-deoxy-d-mannooctulosonic acid - 3-OH-16:0 3-hydroxypalmitic acid - nir^- nitrite reductase negative - nir⁺ nitrite reductase positive     

Chemical composition of lipopolysaccharides from Legionella bozemanii and Legionella longbeachae

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.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     

Modification of cell membrane lipids inMicrococcus lysodeikticus induced by pantoyl lactone

Summary Growth ofMicroccoccus lysodeikticus in the presence of pantoyl lactone brings about both qualitative and quantitative changes in cell membrane lipids. Significant amounts of the two major phospholipids (phosphatidylglycerol and diphosphatidylglycerol) are converted to lyso forms; the largest conversion occurs in the phosphatidylglycerol. In addition, amounts of several phospholipid fatty acids are changed. Physical alteration of the cell membrane can be demonstrated using differential scanning calorimetry. Although growth and transport are significantly inhibited when pantoyl lactone is present, cells possessing altered cell membrane phospholipids and phospholipid fatty acids, brought about by growth in the presence of pantoyl lactone, transportd-alanine,l-glutamic andl-aspartic acid normally when washed free of the pantoyl lactone.     

Characterization of a novel lipopolysaccharide biosurfactant fromKlebsiella oxitoca

The chemical, physical, and emulsifying properties of BSF-1, which is an extracellular lipopolysaccharide biosurfactant produced byKlebsiella oxytoca strain BSF-1, were studied. BSF-1 was found to be composed mainly of carbohydrate and fatty acids. The average molecular weight was 1,700–2,000 kDa. The polysaccharide fraction containedl-rhamnose,d-galactose,d-glucose, andd-glucuronic acid at a molar ratio of 3∶1∶1∶1. The fatty acid content was 1.1% (w/w) and consisted mainly of palmitic acid (C16∶0), 3-hydroxylauric acid (3-OH-C12∶0), and lauric acid (C12∶0). In terms of thermal properties, BSF-1 was revealed to have inter- and intra-molecular hydrogen bonds. The hydrodynamic volume (intrinsic viscosity) of BSF-1 was 22.8 dL/g. BSF-1 could be maintained as a stable emulsion for 48 h through a low-level reduction in surface tension. The optimal emulsification temperature was 30°C. Emulsification by BSF-1 was efficient at both acidic and neutral pH values.     

Chemical composition of the lipopolysaccharides of Ectothiorhodospira shaposhnikovii,Ectothiorhodospira mobilis,and Ectothiorhodospira halophila

Lipopolysaccharides were isolated from the moderate halophilic Ectothiorhodospira shaposhnikovii slight to and Ectothiorhodospira mobilis and from the extremely halophilic Ectothiorhodospira halophila by the hot phenol-water and purified by the phenol-chloroform-petroleum ether methods. The isolated lipopolysaccharides of all three species contained 3-deoxy-d-manno-octulosonic acid and d-glycero-d-mannoheptose indicating the existence of a core. They contained additionally glucose and uronic acids (E. shaposhnikovii and E. mobilis) or glucose, uronic acids and threonine (E. halophila). Sodium deoxycholate gel-electrophoresis of the three lipopolysaccharides, each showing only one major band, indicated R-type character of the lipopolysaccharides of the three Ectothiorhodospira species.The lipid A fractions of the lipopolysaccharides from E. shaposhnikovii and E. mobilis represented phosphorylated mixed lipid A types with both 2,3-diamino-2,3-dideoxy-d-glucose and d-glucosamine. The lipid A from E. halophila contained also phosphate and 2,3-diamino-2,3-dideoxy-d-glucose but only traces of d-glucosamine, which would indicated lipid A_DAG. The fatty acid spectra were characterized by amide-bound 3-OH-10:0 and 3-OH-12:0 (E. shaposhnikovii), 3-OH-10:0 (E. mobilis), or 3-OH-10:0,3-OH-14:0, and 3-oxo-14-0 (E. halophila). The predominant ester-bound fatty acids were 14:0 and 16:0 (E. shaposhnikovii and E. mobilis), or 12:0 and 14:1 (E. halophila).Abbreviations DAG 2,3-diamino-2,3-dideoxy-d-glucose - Kdo 3-deoxy-d-manno-octulosonic acid - GlcA glucuronic acid - GalA galacturonic acid - GC-MS combined gas liquid chromatographymass spectrometry - GlcN Glucosamine - DOC sodium deoxycholate - LPS lipopolysaccharide - PAGE polyacrylamide gel electrophoresis - PCP phenol-chloroform-petroleum ether     

Effect of carbon source on growth temperature and fatty-acid composition in Thermomonospora curvata

The growth-temperature range of the actinomycete, Thermomonospora curvata, was influenced by the nature of the soluble carbon sources used, which were derived from cellulose, pectin, starch and xylan. This thermophile had the broadest (38 to 65°C) and narrowest (42 to 59°C) temperature range during growth on cellobiose (from cellulose) and 4-deoxy-Lxxx-threo-t-hexoseulose uronic acid (from pectin), respectively. This substrate-temperature interaction was accompanied by changes in cellular fatty acids: uronic-acid-grown cells had relatively low amounts of branched chain fatty acids (particularly iso-16:0) and high amounts of monounsaturated fatty acids (particularly cis-18:1) compared with cells grown on any other substrate. Moreover, uronic-acid-grown cells could not respond to increased growth temperature by altering the ratio of branched chain fatty acids to straight chain fatty acids.F.J. Stutzenberger is with the Department of Microbiology, Clemson University, Clemson, SC 29634-1909, USA; T.C. Jenkins is with the Department of Animal, Dairy and Veterinary Sciences at the same university.     

The structure of the lipid A component of Sphaerotilus natans

The lipopolysaccharide of Sphaerotilus natans afforded a ladder-like pattern of bands in sodium deoxycholate-polyacrylamide gel electrophoresis, indicating the presence of a S-form lipopolysaccharide. The chemical analysis showed neutral sugars (rhamnose, glucose, l-glycero-d-manno-heptose), 3-deoxy-octulosonic acid (Kdo), amino compounds (glucosamine, glucosamine phosphate, ethanolamine and ethanolamine phosphate), and phosphorus. The lipid A fraction contained saturated and unsaturated capric, lauric, and myristic acids, and 3-hydroxy capric acid (3-OH-10:0). Its chemical structure was consisting of a glucosamine disaccharide, glycosidically substituted by a phosphomonoester, and substituted at C-4 by a pyrophosphodiester esterified with ethanolamine. The amino groups of both glucosamines are acylated by 3-hydroxy capric acids and these in turn are substituted by saturated and unsaturated capric, lauric, and myristic acids. Hydroxyl groups of the backbone disaccharide at C-3 and C-3 were also esterified by 3-hydroxy capric acid, those at C-4 and C-6 were unsubstituted. The latter provides the attachment site for Kdo.Abbreviations Kdo 3-deoxy-d-manno-octulosonic acid - 3-OH-10:0 3-hydroxy capric acid - DOC-PAGE deoxycholate-polyacrylamide gel electrophoresis - GC-MS gas chromatography/mass spectrometry - LD-MS laser desorption mass spectrometry - LPS lipopolysaccharide - PS polysaccharide     

Characterization of the lipopolysaccharides from eight strains of the cyanobacterium Synechococcus

Lipopolysaccharides have been isolated from eight strains of the unicellular cyanobacterium Synechococcus. Fucose, mannose, galactose, glucose and glucosamine were found in all of the lipopolysaccharides investigated. Additionally, strain-specific sugars are present and permit the chemotyping of lipopolysaccharide. Chemotype I, comprising three strains with a high G+C content of DNA (71-66 mol%), is characterized by a high rhamnose portion and by 3,6-dideoxy-d-arabino-hexose (tyvelose). Chemotype III, represented by three strains with a low G+C content of DNA (55-48 mol%), contains a mannose-polymer with small amounts of 3-O-methyl-mannose, 4-O-methyl-mannose, 2-keto-3-deoxyoctonate and mannosamine. Lipopolysaccharides of the two strains of chemotype II contain 2,3,4-tri-O-methyl-arabinose.Lipid A is difficult to split off from the polysaccharide moiety, but is present in all lipopolysaccharides from the Synechococcus strains. The presence of Lipid A is supported by the finding of -hydroxy fatty acids, predominantly -hydroxypalmitic acid. The distribution of branched -hydroxy fatty acids, detected in small amounts, parallels chemotyping of lipopolysaccharide based on the sugar composition. The phosphorus content of the lipopolysaccharides is low.The pyrogenicity of lipopolysaccharides from two strains is low. Synechococcus lipopolysaccharides have little reactivity in antisera raised in rabbits against homologous cells. As far as tested they do not migrate in immunoelectrophoresis. This confirms the neutral character or low negative charge of Synechococcus lipopolysaccharides.Dedicated to Professor Otto Kandler on occasion of his 60th birthday     

Structural characterization of lipid A component of Erwinia carotovora lipopolysaccharide

The chemical structure of the lipid A component of lipopolysaccharide excreted into the liquid medium by the plant pathogenic enterobacterium Erwinia carotovora FERM P-7576 was characterized. It consists of a -1, 6-linked glucosamine disaccharide which carries ester-and amide-bound fatty acids and phosphate similar to the lipid A from other gram-negative bacteria. The lipid A preparation was not uniform in the number and composition of the fatty acids linked to the disaccharide. Four prominent lipids A were involved, they were composed of five to seven residues of fatty acid. Among them the major component was hexa-acyl lipid A, in which the hydroxyl group at position 3 and the amino group of the non-reducing glucosamine unit carry 3-dodecanoyl-oxytetradecanoyl residues. Positions 2 and 3 of the reducing glucosamine unit were substituted by 3-hydroxytetradecanoic acid. In the hepta-acyl lipid A, an additional hexadecanoic acid was linked to the hydroxyl group of the 3-hydroxytetradecanoyl residue at position 2 of the hexa-acyl lipid A. Two penta-acyl lipids A were the homologs of the hexa-acyl lipid A with decreasing acylation. Dodecanoic acid was missing from one, and 3-hydroxytetradecanoic acid from another. 3-Dodecanoyloxytetradecanoyl residue at position 3 differentiates E. carotovora lipid A from that of other gram-negative bacteria.Abbreviations LPS lipopolysaccharide - GlcN glucosamine - KDO 3-deoxy-d-manno-octulosonic acid - FAB-MS fast atom bombardment mass spectrometry - u atomic mass unit     

Sucrose phosphorylases catalyze transglycosylation reactions on carboxylic acid compounds

Two sucrose phosphorylases were employed for glycosylation of carboxylic acid compounds. Streptococcus mutans sucrose phosphorylase showed remarkable transglycosylating activity, especially under acidic conditions. Leuconostoc mesenteroides sucrose phosphorylase exhibited very weak transglycosylating activity. Three main products were detected from the reaction mixture using benzoic acid and sucrose as an acceptor and a donor molecule, respectively. These compounds were identified as 1-O-benzoyl α-d-glucopyranoside, 2-O-benzoyl α-d-glucopyranose, and 2-O-benzoyl β-d-glucopyranose by 1D-and 2D-NMR analyses of the isolated products and their acetylated products. Time-course analyses proved that 1-O-benzoyl α-d-glucopyranoside was initially produced by the transglycosylation reaction of the enzyme. 2-O-Benzoyl α-d-glucopyranose and 2-O-benzoyl β-d-glucopyranose were produced from 1-O-benzoyl α-d-glucopyranoside by intramolecular acyl migration reaction. S. mutans sucrose phosphorylase showed broad acceptor-specificity. This sucrose phosphorylase catalyzed transglycosylation to various carboxylic compounds such as short-chain fatty acids, hydroxy acids, dicarboxylic acids, and phenolic carboxylic acids. 1-O-Acetyl α-d-glucopyranoside was also enzymatically synthesized by transglucosylation reaction of the enzyme. The sensory test of acetic acid and the glucosides revealed that the sour taste of acetic acid glucosides was significantly lower than that of acetic acid.     

Cell wall and sheath constituents of the cyanobacterium Gloeobacter violaceus

Sheaths isolated from Gloeobacter violaceus were found to be composed of a major polysaccharide moiety (glucose, galactose, rhamnose, mannose, arabinose), a protein moiety, and negatively charged components (glucuronic acids, phosphate, sulfate). Outer membrane polypeptide patterns were dominated by two major peptidoglycan-associated proteins (M_r 62,000 and 53,000). Lipopolysaccharide constituents were glucosamine, 3-hydroxy fatty acids (3-OH-14:0, anteiso-3-OH-15:0, 3-OH-16:0, 3-OH-18:0), carbohydrates, and phosphate. A1-type peptidoglycan and non-peptidoglycan components (mannosamine, glucose, mannose, and glucosamine) indicated the presence of a peptidoglycan-polysaccharide complex in the cell walls of Gloeobacter violaceus.Abbreviations A₂pm diaminopimelic acid - ATCC American Type Culture Collection - CE cell envelope - CM cytoplasmic membrane - CW cell wall - dOcla 3-deoxy-d-manno-2-octulosonic acid - GalN galactosamine - GlcN glucosamine - GlcUA glucuronic acid - HF hydrofluoric acid - LPS lipopolysaccharide - ManN mannosamine - M relative molecular mass - MurN muramic acid - MurN-6-P muramic acid-6-phosphate - OMe O-methyl - PAGE polyacrylamide gel electrophoresis - PCC Pasteur Culture Collection - SDS sodium dodecyl sulfate - SH sheath     

“Oil hyphae” of endolithic lichens and their fatty acid composition

Summary The structure of medullary oil hyphae of twelve endolithic lichen species, belonging to different taxa and colonizing different habitats, was examined by light and electron microscopy. The chemical composition of lipids isolated from the oil hyphae and from two corresponding mycobionts grown in culture was determined.The oil hyphae of the various species appeared in different forms and contained large amounts of lipid in the form of oil globules. The hyphae of mycobionts isolated from two of the endoliths and grown in culture also contained large amounts of lipids. Triacylglycerol was the predominant lipid component in all the organisms examined. Hexadecanoic acid was the main saturated fatty acid; octadecenoic acid and octadecdienoic acid the predominant unsaturated fatty acids. Tetradecanoic, hexadecenoic, octadecanoic and octadectrienoic acids were also detected.The fatty acid distribution pattern appeared unaffected by the nature of substrate and climatic conditions. There is a certain similarity in the fatty acid composition in related species.This work forms part of a dissertation to be submitted by E.Kushnir to the Department of Botany, Tel-Aviv University, in partial fulfillment of the requirements for the degree of Doctor of Philosophy.     

Limitations in the use of 3-hydroxy fatty acid analysis to determine endotoxin in mammalian samples

3-Hydroxy fatty acids (3-OH FAs) of 10–18-carbon chain lengths are constituents of the lipopolysaccharide of Gram-negative bacteria. These acids are used as chemical markers for determining endotoxin in environmental samples. The present communication addresses the question whether this type of analysis also would be applicable to mammalian samples. Low levels (6.1±1.6–94.0±23.2 pmol/ml) of the studied 3-OH FAs were detected in blood from both conventional and germ-free rats. The levels were considerably higher (0.0–1.06±0.17 nmol/mg) in livers. The amounts of the 3-OH FAs did not differ between the two groups of rats. All analyses were made by gas chromatography–tandem mass spectrometry (GC–MSMS) for unequivocal identification. The results illustrate a limitation in using 3-OH FA analysis to determine endotoxin in mammalian samples since these acids may represent not only endotoxin but also products from mammalian mitochondrial fatty acid β-oxidation.     

Fatty acid synthesis inTrichophyton rubrum

Analysis of the fatty acid content ofT. rubrum showed that the organism is capable of synthesizing a variety of saturated and unsaturated long-chain fatty acids (Kostiw, Vicher &Lyon, 1966). The present study was undertaken to determine the mechanism of synthesis of these fatty acids. Experimental data point to the presence of two mechanisms of fatty acid synthesis inT. rubrum:de novo synthesis and chain elongation. The presence of both mechanisms is suggested by the nature of the enzyme preparation, by the cofactor requirements for either pathway, and by the reaction products.University of Illinois at the Medical Center, and Chicago Medical School, Chicago, Illinois. Presented in part at the International Society of Mycology Meeting, New Orleans, Louisiana, August 1967.     

Chemical composition of a lipopolysaccharide from Legionella pneumophila

Lipopolysaccharide isolated from Legionella pneumophila (Phil. 1) was examined for chemical composition. The polysaccharide split off by mild acid hydrolysis contained rhamnose, mannose, glucose, quinovosamine, glucosamine and 2-keto-3-deoxyoctonate, in molar proportions 1.6:1.8:1.0:1.5:4.1:2.7. Heptoses were absent and glucose was probably mainly phosphorylated. The carbohydrate backbone of the lipid A part consisted of glucosamine, quinovosamine and glycerol, in the molar ratios 3.9:1.0:3.4, with glycerol as a phosphorylated moiety. A complex fatty acid substitution pattern comprising eight O-ester-linked, exclusively nonhydroxylated acids, and nineteen amide-linked, exclusively 3-hydroxylated acids was revealed. Both straight- and branched (iso and anteiso) carbon chains occurred. The major hydroxy fatty acid was 3-hydroxy-12-methyltridecanoic acid and six others were of a chain-length above 20 carbon atoms, with 3-hydroxy-20-methyldocosanoic acid as the longest. Two dihydroxy fatty acids, 2,3-dihydroxy-12-methyltridecanoic and 2,3-dihydroxytetradecanoic acids, were also detected. These results suggest that L. pneumophila contains a rather complex and unusual lipopolysaccharide structure of considerable biological and chemotaxonomic interest.Abbreviations LPS lipopolysaccharide - PS polysaccharide - KDO 2-keto-3-deoxy-octonate - GC gas chromatography - GC-MS gas chromatograph-mass spectrometer combined instrument - CI chemical ionization - EI electron impact - HF hydrofluoric acid - TFA trifluoroacetyl - TMS trimethylsilyl     

Lipopolysaccharides of Thiobacillus species containing lipid A with 2,3-diamino-2,3-dideoxyglucose

Lipopolysaccharides were isolated from two strains of Thiobacillus ferrooxidans and one strain each of Thiobacillus thiooxidans, Thiobacillus novellus and Thiobacillus sp. IFO 14570. Neutral sugars, 2-keto-3-deoxyoctonate, fatty acids and the rare 2,3-diamino-2,3-dideoxyglucose were detected in all lipopolysaccharides. Lipopolysaccharides of both T. ferrooxidans strains contained l-glycero-d-manno-heptose, whereas that of T. thiooxidans contained both l-glycero-d-manno-heptose and d-glycero-d-manno-heptose. On the other hand, heptoses were absent in lipopolysaccharides of T. novellus and Thiobacillus sp. IFO 14570. Lipid A of T. ferrooxidans and T. thiooxidans contained both glucosamine and 2,3-diamino-2,3-dideoxyglucose, in contrast, lipid A of T. novellus and the Thiobacillus sp. IFO 14570 most likely contain only 2,3-diamino-2,3-dideoxyglucose as backbone sugar. Deoxycholate polyacrylamide gel electrophoresis revealed S-type character for all lipopolysaccharides studied. The significance of the lipopolysaccharide composition for taxonomic and phylogenetic questions with regard to thiobacilli is discussed.Abbreviations DAG 2,3-diamino-2,3-dideoxyglucose - DOC sodium deoxycholate - GC gas-liquid chromatography - GC/MS gas-liquid chromatography/mass spectrometry - d,d-Heptose d-glycero-d-manno-heptose - l,d-Heptose l-glycero-d-manno-heptose - KDO 2-keto-3-deoxyoctonate - LPS lipopolysaccharide - 3-OH-14:0 3-hydroxy-tetradecanoic acid - PAGE polyacrylamide gel electrophoresis - PCP phenol-chloroform-petroleum ether