Fatty Acid and Polar Lipid Composition in the Classification of Kurthia

Strains of Kurthia zopfii were degraded by acid methanolysis and the non-hydroxylated fatty acid esters so released were examined by gas liquid chromatography. The major fatty acid types were straight-chain, anteiso - and iso -methyl branched-chain acids. Monounsaturated fatty acids were not detected. The major fatty acid in five of the six strains examined consisted of 12-methyltetra-decanoic ( anteiso -C₁₅) acid. The other strain possessed major amounts of both 13-methyltetradecanoic ( iso -C₁₅) and anteiso -C₁₅ acids. Polar lipids of all the strains were examined by two-dimensional thin-layer chromatography. All possessed a very simple polar lipid composition consisting of diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine.     

Temperature-dependent changes in phospholipid and fatty acid composition and membrane lipid fluidity of Yersinia enterocolitica

Temperature-dependent compositional changes of phospholipids and their fatty acids were analysed in Yersinia enterocolitica grown at 5°, 25° and 37°C. The relative amounts of the four phospholipids, phosphatidylethanolamine (75–78%), phosphatidylglycerol (10–11%), cardiolipin (<7%) and lysophosphatidylethanolamine (<5%), were essentially the same at all growth temperatures. The degree of fatty acid unsaturation of the four phospholipids increased with decrease in growth temperature, mainly due to an increase of C_16:1 and C_18:1 and a corresponding decrease of C_16;0, C_18:0 and cyclo C_17:0. An electron spin resonance spectroscopic study of the membrane lipids showed that membrane lipid fluidity was enhanced by decreasing the growth temperatures. The changes in fatty acid composition of phospholipids in response to varied temperatures were consistent with the temperature-dependent changes in the membrane lipid fluidity of Y. enterocolitica , and were similar to those reported for other bacteria.     

Effect of non-target cells on the sensitivity of the PCR for Escherichia coli O157 : H7

The fatty acid composition from mycelia of Streptomyces hygroscopicus strains was studied. A significant proportion of C_{18 : 2} was found in cultures. High levels of C_{16 : 0}, iso-C_{16 : 0} and C_{18 : 1} were also detected in all S. hygroscopicus strains. The different representatives of S. hygroscopicus had almost the same proportion of unsaturated fatty acids. Certain shifts in the amount of iso, anteiso and straight-chain fatty acids in some cultures were revealed. This might be explained by the adaptation capability of strains belonging to one species to form a variety of available fatty acids determined by particular cell membrane composition favouring certain antibiotic biosynthesis.     

Fatty acid, isoprenoid quinone and polar lipid composition in the classification of Renibacterium salmoninarum

Twenty-one strains of Renibacterium salmoninarum were degraded by acid methanolysis and the non-hydroxylated fatty acid esters released examined by thin-layer and gas chromatography. The fatty acid profiles were composed almost exclusively of methyl-branched fatty acids with 12-methyltetradecanoic ( anteiso -C₁₅), 13-methyltetradecanoic ( iso -C₁₅) and 14-methylhexadecanoic ( anteiso -C₁₇) as major components. Polar lipids of the test strains were examined by two-dimensional thin-layer chromatography. All of the organisms possessed very characteristic polar lipid patterns consisting of diphosphatidylglycerol, two major and six or seven minor glycolipids, and two unidentified minor phospholipids. In all cases the major menaquinone components consisted of unsaturated menaquinones with nine isoprene units. The lipid data support the integrity of the genus Renibacterium and can be used to separate it from Corynebacterium and from coryneform bacteria which also contain lysine in the wall peptidoglycan.     

Effect of growth temperature and salt concentration on the fatty acid composition of Flavobacterium halmephilum CCM2831

Abstract In this study the fatty acid composition of Flavobacterium halmephilum CCM2831 grown at different temperatures and salt concentrations is reported. At elevated growth temperatures the amount of cellular saturated long-chain acids (C_18:0 and C_20:0) and the branched-chain acid br-C_17:0 increased, and the concentrations of both cyclic acids and the branched chain acid br-C_15:0 decreased. Increasing the salt concentration in the medium resulted in a gradual increase in cellular cyclopropanoic acids and a concomitant decrease in the monounsaturated fatty acid content.     

Fatty acid and lipid compositions of Conidiobolus

The fatty acid composition of the total lipids from two Conidiobolus species was studied by gas—liquid chromatography. The major fatty acids of C. lamprauges were palmitic acid (C_16:0), oleic acid (C_18:1), linolenic acid (C_18:3), and arachidonic acid (C_20:4). For C. eurymitus , myristic acid (C_14:0), C_16:0, and linoleic acid (C_18:2) were the most abundant acids. The fatty acid composition of C. eurymitus was quite different from that of the Conidiobolus species as mentioned in other reports. The lipid composition of the total lipids of C. lamprauges and C. eurymitus was also studied by thinchrography on quartz rods. Triglycerides and phospholipids were the major components in the two Conidiobolus species.     

Abnormality of Long-Chain Fatty Acids in Erythrocyte Membrane Sphingomyelin from Patients with Adrenoleukodystrophy

Abstract: We have devised an analytical method for the determination of fatty acid composition of erythrocyte membrane sphingomyelin by chemical ionization mass spectrometry combined with capillary column gas-liquid chromatography. Fatty acid composition of erythrocyte membrane sphingomyelin from 8 patients with adrenoleukodystrophy (ALD) and 16 healthy controls were examined by this method. The ratio of hexacosanoic acid (C_26.0) to docosanoic acid (C_22:0) in erythrocyte membrane sphingomyelin from ALD patients was 2.6-fold higher than that of the controls. This result suggests that biochemical diagnosis of ALD is possible by the analysis of fatty acid composition of erythrocyte membrane sphingomyelin. Furthermore, it demonstrates that biochemical abnormality in ALD is the generalized abnormal metabolism of very long-chain saturated fatty acids.     

FATTY ACID COMPOSITION OF CEREBROSIDES, SULPHATIDES AND CERAMIDES IN MURINE LEUCODYSTROPHY: THE QUAKING MUTANT

Abstract— The fatty acid composition of cerebrosides, sulphatides and ceramides has been determined at 20 days postpartum in the brains of Quaking mutant mice and of littermate controls. There was a significant deficit in the proportion of long-chain fatty acids (C₂₂-C₂₄) affecting both normal and a-hydroxy fatty acids of the cerebrosides. The proportion of normal but not the a-hydroxy long-chain fatty acids of the sulphatides was also decreased. Striking and disproportionate deficits of the C_24:1 and C_{24 h:1} fatty acids of cerebrosides, sulphatides and ceramides characterized the brain of the Quaking mutant, and an increased proportion of C_{23 h:O} fatty acid was found in the cerebrosides and sulphatides of the brain of this mutant. We compared these data with findings on the Jimpy mutant which has been examined by the same techniques. The deficiency of long-chain fatty acids which was found in the cerebrosides and sulphatides of both mutants was less extensive but more selective in the Quaking mutant.     

Choline Uptake in Cultured Human Y79 Retinoblastoma Cells: Effect of Polyunsaturated Fatty Acid Compositional Modifications

Abstract: Choline uptake in Y79 human retinoblastoma cells occurs through two kinetically distinguishable processes. The high-affinity system shows little sodium or energy dependence, and it does not appear to be linked to acetyl CoA: choline O -acetyltransferase. When the cells are grown in a culture medium containing 10% fetal bovine serum, the high-affinity system has a K' _m= 2.16 ± 0.13 μ m and V' _max= 27.0 ± 2.9 pmol min⁻¹ mg⁻¹, whereas the low-affinity system has K' _m= 20.4 ± 1.3 μ m and V' _max= 402 ± 49 pmol min⁻¹ mg⁻¹. Under these conditions, the polyunsaturated fatty acid content of the cell membranes is relatively low. When the polyunsaturated fatty acid content of the microsomal membrane fraction was increased by supplementing the culture medium with linolenic or docosahexaenoic acids (n-3 polyunsaturated fatty acids) or arachidonic acid (n-6 polyunsaturated fatty acid), the K' ^m of the high-affinity choline transport system was reduced by 40–60%. The V' _max also was reduced by 20–40%. Supplementation with oleic acid, the most prevalent monounsaturated fatty acid, did not affect either kinetic parameter. The results suggest that one functional effect of the high unsaturated fatty acid content of neural cell membranes is to facilitate the capacity of the high-affinity choline uptake system to transport low concentrations of choline. This effect appears to be specific for polyunsaturated fatty acids but not for a single type, for it is produced by members of both the n-3 and n-6 classes of polyunsaturated fatty acids.     

Wax Ester Production by the Marine Cryptomonad Chroomonas salina Grown Photoheterotrophically on Glycerol

SYNOPSIS. An age-autolyzed culture of Chroomonas salina , grown under cool-white light with glycerol, produced waxy lipid constituting about 44% of total matter harvested. This lipid was composed of 87% wax ester, 9% triglyceride, 3% polar lipid and 1% hydrocarbon. The major wax ester species were identified by total carbon number as C₂₆(28%), C₂₈(35%), C₃₀(15%). The main fatty acid components of the wax esters were 12:0 (39%), 14:0 (30%), 16:0 (14%), while the main alcohols were 14:0 (53%) and 16:0 (40%). The hydrocarbon fraction showed saturated paraffins ranging from C₁₇ to C₃₃, with odd-numbered chain components predominating. No polyunsaturated components were detected in the wax ester or hydrocarbon fractions. This is the first record of wax ester production by a cryptomonad or a marine phytoplankter.     

Nisin induces changes in membrane fatty acid composition of Listeria monocytogenes nisin-resistant strains at 10°C and 30°C

Listeria monocytogenes isolates resistant to 10⁵ IU ml^-1 nisin were obtained at 30°C (NR30) and at 10°C (NR10). Nisin prolonged the lag phase of isolate NR30 at 10°C. Isolates NR30 and NR10 did not produce a nisinase. Protoplasts of isolate NR30 were unaffected by exposure to nisin. The fatty acid composition from the wild-type strain and NR isolates was determined. As expected, temperature-induced differences in the C15/C17 fatty acid ratios were found. Growth of the NR strains in the presence of nisin resulted in significantly different C15/C17 ratios and a significant increase in the percentage of C16:0, C16: 1, C18:0 and C18: 1 fatty acids at 10°C and 30°C. Both the NR10 and NR30 isolates had similar growth rates at low temperatures, but these were slower than the wild-type strain. These results indicate that 'nisin resistance'is an environmentally defined phenotype and that nisin induces changes in the fatty acid composition of the membrane in L. monocytogenes nisin-resistant isolates regardless of the growth temperature.     

The Effect of Temperature on the Fatty Acid Composition of Tetrahymena pyriformis WH-14

SYNOPSIS. A reduction in the growth temperature of Tetrahymena pyriformis strain WH-14 from 35 C to 15 C resulted in distinct alterations in the fatty acid composition of the glycerophospholipids. The proportion of normal saturated acids declined from 26 to 19%; palmitoleic acid increased by 6%, and the composition of the polyunsaturated fatty acids increased in 18:2 Δ^6,11(n) and decreased in 18:2 Δ^9,12(n) and 18:3 Δ^6,9,12(n). The unsaturation index (the average number of double bonds/100 molecules) did not change with a shift in temperature.
Two biosynthetic pathways exist in Tetrahymena for the formation of unsaturated fatty acids. The observed changes in fatty acid composition that accompany a lowering of the environmental temperature can be accounted for by a reduction in the accumulation of products of the fatty acid pathway leading to the formation of γ-linolenic acid [16:0(n) → 18:0(n) → 18:1 Δ⁹(n) → 18:2 Δ^9,12(n) → 18:3 Δ^6,9,12(n)] and an increase in the components of the pathway leading to the formation of 18:2 Δ^6,11(n) [16:0(n) → 16:1 Δ⁹(n) → 18:1 Δ¹¹(n) → 18:2 Δ^6,11(n)]. The data suggest that the regulatory mechanism in Tetrahymena differs from that found in some bacteria where a simple substitution of unsaturated fatty acids for saturated fatty acids occurs at low culture temperatures.     

Myelin Gangliosides in Developing Rats: The Influence of Maternal Ethanol Consumption

Abstract: The present study examined myelin gangliosides in the developing offspring of rats that were pair-fed control or ethanol liquid diets prior to and during gestation. Between 17 and 31 days of age, we observed an increase in the proportion of G_M1 in myelin (from 15% to 38% of ganglioside sialic acid) and a decrease in the proportion of G_T1b (from 26% to 4%). G_M4 was detected at all ages examined. Between 17 and 31 days of age, there was an increase in the proportion of N -acetylman-nosamine-derived radioactivity associated with G_M1 (from 16% to 22%) and G_M4 (from 5% to 13%), and a decrease in that associated with G_T1b (from 24% to 4%). Small, but sygnificant (p < 0.05), developmentally related differences were found in G_D2 and G_D3. Detection of G_M4 in myelin of young rats in the present study appears to depend on the use of nonpartitioning methods of ganglioside extraction. Although the distribution of myelin gangliosides and radioactivity was near-normal in ethanol-treated pups, there was a consistent decrease in the proportion and radioactivity associated with the major myelin ganglioside, G_M1.     

Carbon isotope-labelling experiments indicate that ladderane lipids of anammox bacteria are synthesized by a previously undescribed, novel pathway

Ladderane lipids are unusual membrane lipids of bacteria that anaerobically oxidize ammonium to dinitrogen gas (anammox). Ladderane lipids contain linearly concatenated cyclobutane rings for which the pathway of biosynthesis is currently unknown. To investigate the possible biosynthetic routes of these lipids, 2-¹³C-labelled acetate was added to a culture of the anammox bacterium Candidatus Brocadia fulgida. Labelling patterns obtained by high-field ¹³C nuclear magnetic resonance spectroscopy of isolated lipids indicated that C . Brocadia fulgida synthesizes C_16:0 and iso C_16:0 fatty acids according to the known pathway of type II fatty acid biosynthesis. The ¹³C-labelling pattern of the C₈ alkyl chain of the C₂₀ [3] ladderane monoether also indicated the use of this route. However, carbon atoms in the cyclobutane rings and the cyclohexane ring were nonspecifically labelled and did not correspond to known patterns of fatty acid synthesis. Taken together, our results indicate that it is unlikely that ladderane lipids are formed from the cyclization of polyunsaturated fatty acids as hypothesized previously and suggest an alternative, although as yet unknown, pathway of biosynthesis.     

Identification of the Palmitoylation Site in Rat Myelin P0 Glycoprotein

Abstract: P₀ glycoprotein, the major protein of PNS myelin, contains approximately 1 mol of covalently bound long-chain fatty acids. To determine the chemical nature of the fatty acid-protein linkage, P₀ was labeled in rat sciatic nerve slices with [³H]palmitic acid and subsequently treated with various reagents. The protein-bound palmi-tate was released by incubation with the reducing agents dithiothreitol and 2-mercaptoethanol, and with 1 M hydrox-ylamine at pH 7.5. In addition, P₀ was deacylated by treatment with 10 m M NaBH₄ with the concomitant production of [³H]hexadecanol, indicating that the fatty acid is bound in a thioester linkage. This conclusion was supported further by the fact that deacylation with hydroxylamine generated free thiol groups, which were titrated with [¹⁴C]-iodoacetamide. To identify the cysteine residue involved in the thioester linkage, [¹⁴C]carboxyamidomethylated P₀was digested with trypsin and the resulting peptides analyzed by reversed-phase HPLC. Identification of the radioactive protein fragments by amino acid analysis and amino-terminal peptide sequencing revealed that Cys¹⁵³ in rat P₀ glycoprotein is the acylation site. The acylated cysteine is located at the junction of the putative transmem-brane and cytoplasmic domains. This residue is also present in the P₀ glycoprotein of other species, including human, bovine, mice, and chicken.     

Cellular fatty acid composition in moderately halophilic Gram-negative rods

The fatty acid compositions for 40 strains of moderately halophilic Gram-negative rods were determined by gas-liquid chromatography. The strains studied were included in the genera: Vibrio, Deleya, Alteromonas, Chromobacterium, Flavobac-terium and Pseudomonas. Although there were quantitative differences all strains showed more or less similar spectra of fatty acids ranging from C₁₂ to C₂₀ chain. The major fatty acid species were C_16:0, C_16:1 and C_18:1. Most striking was the predominance of the C_18:1 component, the major fatty acid in extracts of 29 of the 40 strains.     

Description of Oxalicibacterium horti sp. nov. and Oxalicibacterium faecigallinarum sp. nov., new aerobic, yellow-pigmented, oxalotrophic bacteria

Three strains of aerobic, Gram-negative, rod-shaped, non-spore-forming, yellow-pigmented bacteria (OD1^T, YOx^T and NS13), which were isolated in previous studies by enrichment in a mineral medium with potassium oxalate as the sole carbon source, were characterized. On the basis of 16S rRNA gene sequence similarity, strains OD1^T, YOx^T and NS13 belong to the Betaproteobacteria , most closely related to Oxalicibacterium flavum TA17^T (97.2–99.7% sequence similarity). The major whole-cell fatty acids were C_16:0, C_16:1ω7c and C_17:0 cyclo. The results of DNA–DNA hybridization and physiological and biochemical tests allowed genotypic and phenotypic differentiation of strains OD1^T and YOx^T from O. flavum TA17^T and from each other. Therefore, it is concluded that the strains OD1^T and YOx^T represent novel species within the genus Oxalicibacterium , for which the names Oxalicibacterium horti sp. nov. (type strain OD1^T=DSM 21640^T=NBRC 13594^T) and Oxalicibacterium faecigallinarum sp. nov. (type strain YOx^T=DSM 21641^T=CCM 2767^T) are proposed.     

Free Fatty Acid Composition of Human and Rat Peripheral Nerve

Abstract: The free fatty acid (FFA) composition of peripheral nerve resembles that of erythrocytes but the composition of both is different from that of brain and other tissues. Approximately 75% of FFAs of nerve and erythrocytes are saturated and <5% are polyunsaturated whereas in brain and other tissues, 30-45% of FFAs are saturated and 25-50% are polyunsaturated. Approximately 10-15% of the total FFA of nerve have very long chain lengths [C₂₄, C₂₆, C₂₈, and C₃₀]. The presence of these very long-chain FFAs in endoneurium cannot be accounted for by the retention of erythrocytes or by lipid degradation. During Wallerian degeneration a significant increase of 18:1, associated with a decrease of saturated FFAs, was found in rat sciatic endoneurium, but normal values were approached when fiber regeneration was well under way. The FFA composition with chain length ≥C₂₆ were not, however, significantly altered with degeneration or repair of nerves. The metabolic significance of this striking difference between nerve and brain FFA composition is unknown but may reflect different functional properties.     

Multiple controls for the variability of hydrogen isotopic compositions in higher plant n-alkanes from modern ecosystems

We performed a global scale analysis of available leaf wax n -alkane δ D data compiled from our new results, as well as from the literature and expressed as average values of D/H ratios from three common lipids of n -alkanes with odd carbon numbers ( n -C₂₇, n -C₂₉, and n -C₃₁) from living higher plants. Our results clearly indicate multiple controls of hydrogen isotope composition and its variability in plants leaf wax. (1) At the global scale, precipitation δ D values play a dominating factor that exercises the first order of control for hydrogen isotopic compositions in plant leaf wax. The hydrogen isotopic composition of plant leaf wax tracks the decreasing trend of precipitation δ D with increasing latitude. (2) Because of different water acquisition systems, plant life form influences the hydrogen isotopic composition of leaf wax n -alkanes with woody plants and grasses having different responses to the change of global precipitation δ D. (3) Physiological difference, due to different photosynthesis pathways or different water usage strategies, can leave an imprint on δ D patterns of plant leaf waxes, causing δ D variations among plants using the same source water. While these results better explain the variability of hydrogen isotope composition in leaf wax, they also have important implications for the interpretation of n -alkane δ D data from fossils and ancient sediments.     

LIPID CLASS DISTRIBUTION OF HIGHLY UNSATURATED LONG CHAIN FATTY ACIDS IN MARINE DINOFLAGELLATES

The very long chain highly unsaturated C₂₈ fatty acids, octacosaheptaenoic [28:7(n-6)] and octacosaoctaenoic acid [28:8(n-3)], were found to be associated with phospholipids, obtained by fractionation of total lipid extracts into distinct lipid classes, in 4 and 6, respectively, of 16 examined dinoflagellates. An interfraction comparison of fatty acids associated with phospholipids and glycolipids has also shown that the phospholipid fractions contained the majority (over 75% in 12 of 16 strains) of docosahexaenoic acid [22:6(n-3)] and traces of tetracosanoic acid (24:0). By contrast, the highly unsaturated C₁₈ fatty acids octadecatetraenoic [18:4(n-3)] and octadecapentaenoic acid [18:5(n-3)] were primarily recovered from a chloroplast-associated glycolipid fraction comprised of monogalactosyldiacylglycerol, digalactosyldiacylglycerol, and sulfoquinovosyldiacylglycerol. In 12 of 16 strains, an interfraction comparison showed that over 90% of 18:5(n-3) was found to be associated with glycolipids. These findings indicate that the C₂₈ fatty acids are located and probably synthesized in the cytoplasm or in an organelle other than the chloroplast, possibly with 22:6(n-3) and 24:0 as precursors, whereas the C₁₈ fatty acids 18:4(n-3) and 18:5(n-3) are glycolipid constituents apparently synthesized within the chloroplast. The function(s) of these C₂₈ fatty acids as components of phospholipids in cellular membranes is currently unknown.