Methyleneoctadecanoic Acid Research Articles

Cell envelope phospholipid composition of Burkholderia multivorans.

Burkholderia multivorans causes opportunistic pulmonary infections in cystic fibrosis and immunocompromised patients. The purpose of the present study was to determine the nature of the phospholipids and their fatty acid constituents comprising the cell envelope membranes of strains isolated from three disparate sources. A conventional method for obtaining the readily extractable lipids fraction from bacteria was employed to obtain membrane lipids for thin-layer chromatographic and gas chromatography-mass spectrophotometric analyses. Major fatty acid components of the B. multivorans readily extractable lipid fractions included C(16:0) (palmitic acid), C(16:1) (palmitoleic acid), and C(18:1) (oleic acid), while C(14:0) (myristic acid), ΔC(17:0) (methylene hexadecanoic acid), C(18:0) (stearic acid), and ΔC(19:0) (methylene octadecanoic acid) were present in lesser amounts. Fatty acid composition differed quantitatively among strains with regard to C(16:0), C(16:1), ΔC(17:0), C(18:1), and ΔC(19:0) with the unsaturated:saturated fatty acid ratios being significantly less in a cystic fibrosis type strain than either environmental or chronic granulomatous disease strains. Phospholipids identified in all B. multivorans strains included lyso-phosphatidylethanolamine, phosphatidylglycerol, phosphatidylethanolamine, and diphosphatidylglycerol in similar ratios. These data support the conclusion that the cell envelope phospholipid profiles of disparate B. multivorans strains are similar, while their respective fatty acyl substituent profiles differ quantitatively under identical cultivation conditions.

A shotgun lipidomics approach in Sinorhizobium meliloti as a tool in functional genomics

A shotgun lipidomics approach that allowed the analysis of eight lipid classes directly from crude extracts of the soil bacterium Sinorhizobium meliloti is presented. New MS-MS transitions are reported for the analysis of monomethylphosphatidylethanolamines, dimethylphosphatidylethanolamines, and three bacterial non-phosphorus-containing lipid classes [sulfoquinovosyldiacylglycerols, ornithines, and diacylglyceryl-(N,N,N-trimethyl)-homoserines]. Unique MS-MS transitions allowed the analysis of isomeric species from various lipid classes without chromatography. Analyses required small sample amounts and minimal preparation; thus, this methodology has excellent potential to be used as a screening tool for the analysis of large numbers of samples in functional genomics studies. FA distributions within lipid classes of S. meliloti are described for the first time, and the relative positions of fatty acyl substituents (sn-1, sn-2) in phospholipids are presented. FA distributions in diacylglyceryl-(N,N,N-trimethyl)-homoserines were identical to those of phospholipids, indicating a common biosynthetic origin for these lipids. The method was applied to the analysis of mutants deficient in the PhoB regulator protein. Increased lipid cyclopropanation was observed in PhoB-deficient mutants under P(i) starvation.

Trans fatty acids promote the growth of some Lactobacillus strains

Five Lactobacillus strains (2 L. gasseri, 2 L. plantarum and 1 L. reuteri) were cultured in modified MRS medium containing fatty acids (FAs) instead of Tween 80 for 24 h at 37 degrees C, to learn the effect of saturated and unsaturated FAs on the Lactobacillus growth. Free FAs included palmitic (16:0), palmitoleic (c9-16:1), stearic (18:0), oleic (c9-18:1), elaidic (t9-18:1), cis-vaccenic (c11-18:1), vaccenic (t11-18:1), linoleic (c9, c12-18:2), conjugated linoleic (c9, t11- and t10, c12-18:2), alpha-linolenic (c9, c12, c15-18:3), alpha-eleostearic (c9, t11, t13-18:3), eicosapentaenoic (20:5), and docosahexaenoic (22:6) acids. Among free FAs, oleic acid stimulated the growth of all Lactobacillus strains, whereas palmitoleic acid had almost no affect on the Lactobacillus growth. Saturated FAs such as stearic and palmitic acids inhibited or did not affect the Lactobacillus growth. Polyunsaturated FAs such as alpha-linolenic, eicosapentaenoic and docosahexaenoic acids strongly inhibited the Lactobacillus growth at 7.6 x 10(-4) m. Octadecenoic acids such as oleic, elaidic, cis-vaccenic and vaccenic acids remarkably promoted the growth of L. gasseri, regardless of the different double bond positions and configurations. When oleic or cis-vaccenic acid was incubated with L. gasseri, the FAs was transformed to cyclopropane FAs (methyleneoctadecanoic acids) after incorporation into the cells. On the other hand, trans FAs such as elaidic and vaccenic acids incorporated into the cells were not converted to another FAs. Conjugated linoleic and alpha-eleostearic acids having a trans double bond promoted the Lactobacillus growth. The growth of L. gasseri was also stimulated by trans-rich free FAs from hydrogenated canola and fish oils. These results showed that octadecenoic acid and trans FAs had strong promotion activities for the Lactobacillus growth due to their incorporation into membrane lipids.

Occurrence of the toxin dehydroabietic acid in Salmonella typhimurium

Many strains of Salmonella typhimurium studied in our lab demonstrated marked differences in the pathogenicity for guinea pig, chicken and Hela cells. As a result, a pathogenic strain of S. typhimurium, strain 9SR2, was evaluated for lipophilic components that may be associated with virulence using gas chromatography/mass spectrometry. The hydroxylated fatty acids 2-hydroxytetradecanoic acid (2-OH-14:0) and 3-hydroxytetradecanoic acid (3-OH-14:0) often present in lipid A, a potent endotoxin, were observed as their methyl esters. The cyclic fatty acids methylene-hexadecanoic acid (C17Δ) and methyleneoctadecanoic acid (C19Δ) also were detected. The nephrotoxic and neurotoxic diterpenoid resin acid, dehydroabietic acid, was observed for the first time from S. typhimurium in both the total lipid and diglyceride fractions and determined as its methyl ester at m/ z 314.2246. Due to its previously established toxicity, dehydroabietic acid may be a factor associated with virulence of S. typhimurium.

The presence of 11-methyloctadec-11-enoic acid in the extractable lipids of Pseudomonas vesicularis

The polar lipids of Pseudomonas vesicularis do not contain a methyleneoctadecanoic acid but instead the isomeric 11-methyloctadec-11-enoic acid. The presence of the latter acid distinguishes the lipids of P. vesicularis from those of P. diminuta, but links them with the lipids of Rhizobium and Caulobacter species.

Polar lipids and fatty acids of Pseudomonas cepacia

The polar lipids and fatty acids produced by the reference strains for seven different O serogroups of Pseudomonas cepacia have been identified. Similar results were obtained for all strains. Contrary to a previous report, the only significant phospholipids in this species are phosphatidylethanolamine and bis(phosphatidyl)glycerol, which contributed 57–83% and 17–43%, respectively, of the total lipid phosphorus. The former lipid was found as two chromatographically distinct fractions. In the less polar fraction and in bis(phosphatidyl)glycerol, the major fatty acids were hexadecanoic acid, cis-9,10-methylenehexadecanoic acid, cis-octadec-11-enoic acid, and cis-11,12-methyleneoctadecanoic acid. In the more polar fraction of phosphatidylethanolamine, the fatty acid in one position is a 2-hydroxy acid, mainly 2-hydroxyhexadecenoic acid, 2-hydroxyhexadecanoic acid, 2-hydroxyoctadecenoic acid, or 2-hydroxymethyleneoctadecanoic acid. Compared with other phospholipids, this fraction of phosphatidylethanolamine was depleted in cis-9,10-methylenehexadecanoic acid. Each strain also produced two ornithine amide lipids. In the major lipid, 3-hydroxyhexadecanoic acid was amide-bound to the α-amino group and was itself probably esterified by a 2-hydroxy acid, mainly 2-hydroxyoctadecanoic acid or the derived cyclopropane acid. In the minor ornithine amide lipid, the ester-bound acids were mainly methyleneoctadecanoic acid and hexadecanoic acid. The unusual lipid profiles of P. cepacia are of chemotaxonomic interest.

Campylobacter pyloridis, gastritis, and peptic ulceration.

Campylobacter pyloridis is a spiral bacterium which was seen by histopathologists several years before it was cultured in 1982 in Perth, Western Australia. It has unique cellular fatty acids, predominantly tetradecanoic acid and cis-11, 12 methylene octadecanoic acid. It also has a unique ultrastructure which is different from that of other campylobacters. C pyloridis possesses a powerful urease enzyme and produces large amounts of extracellular catalase. Both these features may be important virulence factors, allowing it to occupy a protected niche in the stomach below the mucus layer but above the gastric mucosa. Specific lesions are found in the gastric mucosa, and ultrastructural studies show the presence of adherence pedestals identical with those found with enteropathogenic Escherichia coli of the intestine. Histological examination of gastric biopsy tissue has shown that C pyloridis is strongly associated with active chronic gastritis, when polymorphonuclear leucocytes are present, and is not found on normal mucosa except when a biopsy specimen from elsewhere in the stomach shows active chronic gastritis. When patients with symptoms caused by gastritis are identified dual antibacterial treatment, combining the action of bismuth in the stomach with a systemic antibiotic, can eradicate C pyloridis, with remission of symptoms and restoration of normal epithelial morphology. Most peptic ulcers relapse after modern acid reducing treatment, and antibacterial treatment may be beneficial in preventing relapse.

Relationship between cyclopropane synthetase and the formation of cyclopropane fatty acids by Proteus vulgaris grown under various respiratory conditions.

In medium saturated with oxygen, the cyclopropane synthetase (unsaturated-phospholipid methyltransferase; EC 2.1.1.16) of Proteus vulgaris was generally synthesized after the mid-exponential phase of growth. The enzyme could also be induced by rapidly limiting the oxygen supply, or by initiating respiration on nitrate or thiosulphate following an initial period of growth in a highly aerobic environment. In each of these 'step-down' situations the specific activity of cyclopropane synthetase rose to a maximum prior to the stationary phase of growth and subsequently decreased. The cyclopropane fatty acids, methylene hexadecanoic acid and methylene octadecanoic acid accumulated throughout exponential growth following the induction of the enzyme. During a 12 h period in the stationary phase there was little synthesis of either of the fatty acids, despite detectable cyclopropane synthetase activity in the cells, indicating that essentially all the fatty acid synthesis was complete prior to entering the stationary phase. When nitrate was used as a respiratory electron acceptors a twofold increase in octadecenoic acid was observed, giving rise to an increase in methylene octadecanoic acid. This increase in octadecanoic acid was not apparent in mutants unable to respire on nitrate.

The Metabolism of Cyclopropane Fatty Acids by Tetrahymena pyriformis

Abstract Cyclopropane fatty acids are readily metabolized by whole cells of Tetrahymena pyriformis when the fatty acids are presented either as components of intact Escherichia coli cells or as free fatty acids added to the medium. cis-11,12-[methylene-14C]Methyleneoctadecanoic acid was degraded with the production of 14CO2 by whole T. pyriformis cells or a particulate fraction derived from them. In the presence of malonate, [2-14C]acetate was formed. A pathway for degradation of the cyclopropane fatty acids, requiring only minor modification of the usual β oxidation pathway, is proposed.

The effects of temperature on the composition and physical properties of the lipids of Pseudomonas fluorescens.

1. Pseudomonas fluorescens was grown at various temperatures between 5 degrees C and 33 degrees C. The extractable lipids from organisms at various stages of growth and grown at different temperatures were examined. 2. The extractable lipids contained phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, and an ornithine-containing lipid. The relative amounts of these lipids did not vary significantly during growth or with the changes in growth temperature. 3. The major fatty acids were hexadecanoic, hexadecenoic and octadecenoic acids and the cyclopropane acids methylene-hexadecanoic and methylene-octadecanoic acids. The relative amount of unsaturated acids (including cyclopropane acids) did not change significantly during growth, but increased with decreasing temperature. 4. Phosphatidylethanolamines with different degrees of unsaturation and containing different amounts of cyclopropane acids were isolated from organisms grown at 5 degrees C and 22 degrees C and their surface and phase behaviour in water was investigated. Thermodynamic parameters for fusion and monolayer results for cyclopropane and other fatty acids were examined. 5. The surface pressure-area isotherms of phosphatidylethanolamines containing different amounts of unsaturated fatty acids show small differences but the individual isotherms remain essentially unchanged over the temperature range 5-22 degrees C. X-ray-diffraction methods show that the structures (lamellar+hexagonal) formed in water by phosphatidylethanolamine, isolated from organisms grown at 5 degrees C and 22 degrees C, are identical when compared at the respective growth temperatures. This points to a control mechanism of the physical state of the lipids that is sensitive to the operating temperature of the organism. 6. The molecular packing of cyclopropane acids is intermediate between that of the corresponding cis- and trans-monoenoic acids. However, substitution of a cyclopropane acid for a cis-unsaturated acid has insignificant effects on the molecular packing of phospholipids containing these acids.

Phospholipids of Clostridium butyricum. IV. Analysis of the positional isomers of monounsaturated and cyclopropane fatty acids and alk-1′-enyl ethers by capillary column chromatography

The positional isomers of the cyclopropane fatty acids of Clostridium butyricum phospholipids have been analyzed by capillary column gas-liquid chromatography. Greater than 95% of the methylenehexadecanoic acids was the 9,10 isomer. On the other hand, 60-70% of the hexadecenoic acid precursors was the Delta(7) isomer, and the remainder was the Delta(9) isomer. Of the methyleneoctadecanoic acids 75-80% was the 11,12 isomer, with the remainder being the 9,10 isomer. There were approximately equal amounts of the Delta(9)- and Delta(11)-octadecenoic acids in the phospholipids. This study reveals a surprisingly strong specificity of the cyclopropane synthetase for the (n-7) series of monoenoic fatty acids. An analysis by capillary column chromatography of the monoenoic and cyclopropane aldehyde dimethylacetals derived from the plasmalogens (1-alk-1'-enyl-2-acyl-glycero-phosphatides) of C. butyricum revealed the presence of the same positional isomeric mixtures of the 16- and 18-carbon monoenoic residues in approximately the same ratios as were found in the fatty acids. In the formation of the cyclopropane alk-1'-enyl ethers there was also specificity for the (n-7) series, but it was not as strong as that seen in the fatty acids. The ratio of the 7,8 isomer to the 9,10 isomer was higher in the methyl-enehexadecanals than in the corresponding fatty acids. This paper extends the use of Golay capillary columns to the analysis of the positional isomers of plasmalogen aldehydes as their dimethylacetal derivatives.

Some factors affecting cyclopropane acid formation in Escherichia coli.

1. The fatty acid composition of the extractable lipids of Escherichia coli varied with growth conditions. 2. The principal fatty acids were palmitic acid, hexadecenoic acid, octadecenoic acid and the cyclopropane acids, methylenehexadecanoic acid and methyleneoctadecanoic acid. 3. Cyclopropane acid formation from monoenoic acids was increased by acid media, poor oxygen supply, or high growth temperature. 4. Cyclopropane acid formation was decreased by alkaline media, well oxygenated conditions, the presence of citrate, or lack of Mg(2+).

EFFECT OF TEMPERATURE ON THE COMPOSITION OF FATTY ACIDS IN ESCHERICHIA COLI

Marr, Allen G. (University of California, Davis) and John L. Ingraham. Effect of temperature on composition of fatty acids in Escherichia coli. J. Bacteriol. 84:1260-1267. 1962.-Variations in the temperature of growth and in the composition of the medium alter the proportions of individual fatty acids in the lipids of Escherichia coli. As the temperature of growth is lowered, the proportion of unsaturated fatty acids (hexadecenoic and octadecenoic acids) increases. The increase in content of unsaturated acids with a decrease in temperature of growth occurs in both minimal and complex media. Cells harvested in the stationary phase contained large amounts of cyclopropane fatty acids (methylenehexadecanoic and methylene octadecanoic acids) in comparison with cells harvested during exponential growth. Cells grown in a chemostat, limited by the concentration of ammonium salts, show a much higher content of saturated fatty acids (principally palmitic acid) than do cells harvested from an exponentially-growing batch culture in the same medium. Cells grown in a chemostat, limited by the concentration of glucose, show a slightly higher content of unsaturated fatty acids than cells from the corresponding batch culture. The results do not indicate a direct relation between fatty acid composition and minimal growth temperature.

Structure of Malvalic Acid and Some Related C18 and C19 Cyclopropane Fatty Acids

IN recent years an interesting group of fatty acids containing a cyclopropane or a cyclopropene ring has been isolated from natural products or synthesized. This work followed the discovery by Hofmann et al. 1,2 of a biologically active C19H36O2 cyclopropane fatty acid, lactobacillic acid, in the lipides of Lactobacillus arabinosus and L. casei. Four more C19 acids of this type were synthesized3,4—the DL, cis and trans, 9-10 and 11-12, methylene octadecanoic acids. The crystal structures of two of these racemates have been studied5,6, namely, the DL trans 9-10 and DL cis 11-12, and shown to have the characteristic straight- and bent-chain configurations illustrated in the electron density Fourier projections in Figs. 1 and 2. From the similarity in the diffraction data, the other two racemates are believed to be structurally isomorphous7 in the trans and cis series, respectively. Of these, the DL cis 9-10 acid was shown7 to be identical with the dihydrosterculic acid obtained4 by hydrogenation of the sterculic acid8, C19H34O2, from the kernel oil of the tropical tree Sterculia foetida.