Biosynthetic and environmental effects on the stable carbon isotopic compositions of anteiso- (3-methyl) and iso- (2-methyl) alkanes in tobacco leaves

Nicotiana tabacum is the only plant known to synthesise large quantities of anteiso- (3-methyl) alkanes and iso- (2-methyl) alkanes. We investigated the carbon isotope ratios of individual long-chain n-alkanes, anteiso- and iso-alkanes (in the C₂₉-C₃₃ carbon number range) extracted from tobacco grown in chambers under controlled conditions to confirm the pathway used by the tobacco plant to synthesise these particular lipids and to examine whether environmental data are recorded in these compounds. Tobacco was grown under differing temperatures, water availabilities and light intensities in order to control its stable carbon isotope ratios and evaluate isotopic fractionations associated with the synthesis of these particular lipids. The anteiso-alkanes were found to have a predominant even-carbon number distribution (maximising at C₃₂), whereas the iso-alkanes exhibit an odd-carbon number distribution (maximising at C₃₁). Iso-alkanes were relatively more abundant than the anteiso-alkanes and only two anteiso-alkanes (C₃₀ and C₃₂) were observed.The anteiso-alkanes and iso-alkanes were found to be enriched in ¹³C by 2.8-4.3‰ and 0-1.8‰ compared to the n-alkanes, respectively, consistent with different biosynthetic precursors. The assumed precursor for the odd-carbon-numbered iso-alkanes is iso-butyryl-CoA (a C₄ unit derived from valine) followed by subsequent elongation of C₂ units and then decarboxylation. The assumed precursor for even-carbon-numbered anteiso-alkanes is α-methylbutyryl-CoA (a C₅ unit derived from isoleucine) and subsequent elongation by C₂ units followed by decarboxylation. The ratio of carbon atoms derived from α-methylbutyryl-CoA and subsequent C₂ units (from malonyl-CoA) is 1:5 for the biosynthesis of a C₃₀anteiso-alkane. The ratio of carbon atoms derived from iso-butyryl-CoA and subsequent C₂ units (from malonyl-CoA) is 4:25 for the synthesis of a C₂₉iso-alkane. An order of ¹³C depletion n-alkanes > iso-alkanes > anteiso-alkanes is evident from compound specific isotope data. This trend can probably be attributed to the ratio of the two different sources of carbon atoms in the final wax components.Higher water availability generally results in more depleted stable carbon isotope ratios due to maximised discrimination during carboxylation, associated with less diffusional limitation. This was confirmed in the present study by compound specific isotope analyses of iso-alkanes, anteiso-alkanes and n-alkane lipids extracted from the tobacco leaves. Likewise, light intensity has been shown to influence plant bulk δ¹³C in previous studies. The carbon isotope ratios of n-alkanes in tobacco grown under low-light conditions were about 2‰ more depleted in ¹³C than those of lipids extracted from tobacco grown under elevated light conditions. A similar order of difference is observed for the iso-alkanes and anteiso-alkanes (1.8‰ and 1.9‰, respectively). A negligible depletion in carbon isotope ratios was observed for the iso-alkanes and anteiso-alkanes extracted from tobacco grown under elevated temperatures. These results are consistent with the work of Farquhar [Farquhar, G.D., 1980. Carbon isotope discrimination by plants: effects of carbon dioxide concentration and temperature via the ratio of intercellular and atmospheric CO₂ concentrations. In: Pearman, G.I. (Ed.), Carbon Dioxide and Climate: Australian Research. Springer, Berlin, pp. 105-110] where temperature appears to have only a minor effect on plant bulk δ¹³C.     

Lipids of Chlamydomonas reinhardi under different growth conditions

Photo-, mixo- and heterotrophically grown cultures of Chlamydomonas reinhardi (wild type ss and 2 streptomycin-resistant mutants sr₃ and sr₃₅) have been analyzed for lipids and fatty acids. Ether-soluble lipids, chlorophyll, monogalactosyl diglyceride, digalactosyl diglyceride, sulfolipid, phosphatidyl ethanolamine, phosphatidyl choline, phosphatidyl glycerol and the relative amounts of fatty acids in total and individual lipids have been determined. The lipid and fatty acid compositions are very similar in the 3 strains and are not affected by the mutations. Fatty acids belong exclusively to the C₁₆ and C₁₈ series, 16:0, 16:4, 18:1, 18:2, 18:3 (6,9,12) and 18:3 (9,12,15) comprising about 90% of the total. 18:3 (6,9,12) is concentrated in phosphatidyl ethanolamine. In streptomycin-bleached sr₃ cells, ether-soluble lipids increase from 7 to 11% of dry weight on greening, mostly due to synthesis of monogalactosyl diglyceride and chlorophyll. Monogalactosyl diglyceride of bleached cells exhibits the same fatty acid pattern before and after greening.     

Lipid composition of cyanidium

The major lipids in Cyanidium caldarium Geitler are monogalactosyl diglyceride, digalactosyl diglyceride, plant sulfolipid, lecithin, phosphatidyl glycerol, phosphatidyl inositol, and phosphatidyl ethanolamine. Fatty acid composition varies appreciably among the lipids, but the major ones are palmitic acid, oleic acid, linoleic acid, and moderate amounts of stearic acid. Trace amounts of other acids in the C₁₄ to C₂₀ range were also present. Moderate amounts of linolenic acid were found in two strains, but not in a third. The proportion of saturated acid is relatively high in all lipids ranging from about a third in monogalactosyl diglyceride to three-fourths in sulfolipid. This may be a result of the high growth temperature. Lipases forming lysosulfolipid, and lysophosphatidyl glycerol are active in ruptured cells; galactolipid is degraded with loss of both acyl residues. Thus the lipid and fatty acid composition of Cyanidium more closely resembles that of green algae than that of the blue-green algae, although there are differences of possible phylogenetic interest.     

Effect of cultural conditions on the lipid profile of Thiobacillus ferroxidas

The intensitive investigations on the lipid profile of Thiobacillus ferrooxidans at various culture ages suggest some correlations of the lipid constitutents with the membrane-bound iron oxidation system. Phosphatidic acid, phosphatidyl serine and phosphatidyl ethanolamine were the major polar components; hydrocarbon, triglyceride and diglyceride were the main neutral components. Major fatty acids were C_16:0, C_16:1, C_16:3, C_18:1, C_18:3, C_22:1 while C_20:1, C_20:2, C_12:0, C_14:2, C_18:0, C_18:2, C_20:0, C_22:0 were found in trace amounts which also depended upon the phase of the growth. One lipoamino acid was identified as ornithine lipid in the polar fraction. Each and every component varied to some extent at different growth phasesindicating relationship of these lipids to the iron oxidation system of the strain.     

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.     

Fatty acid desaturase 2 (FADS2) but not FADS1 desaturates branched chain and odd chain saturated fatty acids

Branched chain fatty acids (BCFA) and linear chain/normal odd chain fatty acids (n-OCFA) are major fatty acids in human skin lipids, especially sebaceous gland (SG) wax esters. Skin lipids contain variable amounts of monounsaturated BCFA and n-OCFA, in some reports exceeding over 20% of total fatty acids. Fatty acid desaturase 2 (FADS2) codes for a multifunctional enzyme that catalyzes Δ4-, Δ6- and Δ8-desaturation towards ten unsaturated fatty acids but only one saturate, palmitic acid, converting it to 16:1n-10; FADS2 is not active towards 14:0 or 18:0. Here we test the hypothesis that FADS2 also operates on BCFA and n-OCFA. MCF-7 cancer cells stably expressing FADS1 or FADS2 along with empty vector control cells were incubated with anteiso-15:0, iso-16:0, iso-17:0, anteiso-17:0, iso-18:0, or n-17:0. BCFA were Δ6-desaturated by FADS2 as follows: iso-16:0 → iso-6Z-16:1, iso-17:0 → iso-6Z-17:1, anteiso-17:0 → anteiso-6Z-17:1 and iso-18:0 → iso-6Z-18:1. anteiso-15:0 was not desaturated in either FADS1 or FADS2 cells. n-17:0 was converted to both n-6Z-17:1 by FADS2 Δ6-desaturation and n-9Z-17:1 by SCD Δ9-desaturation. We thus establish novel FADS2-coded enzymatic activity towards BCFA and n-OCFA, expanding the number of known FADS2 saturated fatty acid substrates from one to six. Because of the importance of FADS2 in human skin, our results imply that dysfunction in activity of sebaceous FADS2 may play a role in skin abnormalities associated with skin lipids.     

Saturated mono-unsaturated long-chain hydrocarbon profiles of sweet oranges

Three varieties of midseason oranges, viz. Jaffa, Homosassa and Queen, were examined for their saturated and mono-unsaturated hydrocarbon composition in juice sacs. Hydrocarbons were isolated by lipid extraction of the juice sac powders followed by column, thin-layer and AgNO₃-TLC. After hydrogenation, the mono-unsaturated fraction and the saturated fraction were analyzed by GC. In the saturated fraction, the dominant linear hydrocarbon was C₂₃ while C₂₅ predominated in the monoene fraction. iso- And anteiso-branched hydrocarbons comprised between 53 and 63 % of the saturated fraction and only 20–26 % of the monoene fraction. Queen differed from Homosassa and Jaffa in the accumulation of higher percentages of saturated iso- and anteiso-branched hydrocarbons and conversely, showed lower percentages for these branched structures in the monoene fraction. Based on the total relative percentages of the three isomeric hydrocarbons, Homosassa could not be differentiated from Jaffa. The overall profiles for these two oranges, however. showed noticeable differences.     

Effects of phospholipid acyl chain structure on thermotropic phase properties. 3. Phosphatidylcholines with (−), and (±)-anteiso acyl chains

We have synthesized a number of anteiso-branched fatty acids, both as racemates and as pure (?)-stereoisomers, as well as the corresponding di-anteiso-acyl phosphatidylcholines (PCs). The phase transition temperatures (_c of the hydrated PCs have been determined by differential thermal analysis (DTA). Consideration of the observed effects of acyl chain length and terminal see-butyl group configuration on anteiso acid melting points and di-anteiso-acyl PC transition temperatures leads us to the conclusion that the terminal branched portions of anteiso acyl chains interact only weakly with adjacent acyl chains in the hydrated phosphatidylcholine bilayer and probably in the anhydrous fatty acid crystal as well. In agreement with this conclusion, we observe that the DTA heating transition endotherms for hydrated di-anteiso-acyl PCs appear to be considerably less strongly endothermic, and occur at much lower temperatures, than do the major transition endotherms for the di-n-acyl PCs of like carbon number. Our findings generally support the proposal that anteiso acyl lipids tend to fluidize membranes, although they do so less effectively than do cis-unsaturated acyl lipids.     

Salinicoccus halitifaciens sp. nov., a novel bacterium participating in halite formation

Strain JC90^T was isolated from a soda lake in Lonar, India. Strain JC90^T maintains its external pH to 8.5 and participates in halite formation. Based on 16S rRNA gene sequence similarity studies, strain JC90^T was found to belong to the genus Salinicoccus and is most closely related to “Salinicoccus kekensis” K164^T (99.3 %), Salinicoccus alkaliphilus T8^T (98.4 %) and other members of the genus Salinicoccus (<96.5 %). However Strain JC90^T is <36 % related (based on DNA–DNA hybridization) with the type strains of “S. kekensis” K164^T and S. alkaliphilus T8^T. The DNA G+C content of strain JC90^T was determined to be 46 mol %. The cell-wall amino acids were identified as lysine and glycine. Polar lipids were found to include diphosphatidylglycerol, phosphatidylglycerol, phosphatidyl ethanolamine, an unidentified glycolipid and unidentified lipids (L1,2). Major hopanoids of strain JC90^T were determined to be bacterial hopane derivatives (BHD1,2), diplopterol, diploptene and two unidentified hopanoids (UH1,2). The predominant isoprenoid quinone was identified as menaquinone (MK-6). Anteiso-C_15:0 was determined to be the predominant fatty acid and significant proportions of iso-C_14:0, C_14:0, iso-C_15:0, C_16:0, iso-C_16:0, iso-C_17:0, anteiso-C_17:0 and C_18:02OH were also detected. The results of physiological and biochemical tests support the molecular evidence and allowed a clear phenotypic differentiation of strain JC90^T from all other members of the genus Salinicoccus. Strain JC90^T is therefore considered to represent a novel species, for which the name Salinicoccus halitifaciens sp. nov. is proposed. The type strain is JC90^T (=KCTC 13894^T =DSM 25286^T).     

Characteristics of <Emphasis Type="Italic">Pseudomonas aurantiaca</Emphasis> DNA supramolecular complexes at various developmental stages

Differences in viscoelasticity (η) and molecular mass (M) values, as well as in the fatty acid profile of lipids in DNA supramolecular complexes (SC), isolated from Pseudomonas aurantiaca cultures at the exponential and stationary growth phases, were established for the first time. Typical characteristics of DNA SC from actively growing cells were the following: η = 315 ± 15 dl/g, M_DNA = 39 × 10⁶ Da, C_16:0 > C_18:0 > C_18:1 present as basic fatty acids (FA) in a pool of loosely DNA-bound lipids; the tightly DNA-bound lipid fraction consisted of only two acids C_18:0 > C_16:0. Significantly higher values of viscoelasticity η = 779 ± 8 dl/g and M_DNA = 198 × 10⁶ Da were observed for DNA SC of the stationary phase cells; one more FA, C_14:0, was detected in the loosely bound lipid fraction, while lipids tightly bound to DNA contained mainly C_16:0 > C_18:1 > C_18:0 > C_14:0 FA. The content of saturated FA in the DNA-bound lipids in the stationary phase cells was twice as high than in the exponential phase cells. The fraction of tightly bound lipids from the stationary phase cells contained nine times more unsaturated fatty acids than the fraction from proliferating cells. These differences in FA composition of DNA-bound lipids demonstrate the importance of lipids for the structural organization and functioning of genomic DNA during bacterial culture development.     

Differentiation Inducing Phosphatidyl Choline(s) from the Embryos of Rainbow Trout (Salmo gairdneri): Isolation and Structural Elucidation1

Phospholipid which is able to induce the differentiation of some undifferentiated tumour cells was isolated from the rainbow trout (Salmo gairdneri) embryos. As HPLC gave only one peak, the phospholipid seemed to be completely purified. However, the spectra of SIMS and FD-MS have shown that it was a mixture of two molecular species. By the chemical and enzymic studies their structures were determined as C_16:0/C_22:6-and C_18:1/C_22:6-phosphatidyl cholines. The isolated and structurally elucidated phosphatidyl cholines induce haemoglobin synthesis in murine erythroleukemia cells and a rapid decrease in alkaline phosphatase activity in teratocarcinoma cells. Some phosphatidyl cholines which are commercially available have shown no or conspicuously low activities. Recently we isolated another differentiation-inducing substance(s), which proved to be diglyceride(s) containing the completely same fatty acids as those in the above mentioned phosphatidyl cholines, that is, C_16:0/C_22:6-and C_18:1/C_22:6. These results suggest that the lipids containing C_22:6 fatty acid or this fatty acid itself might play an important part in the differentiation and development.     

Combination and positional distribution of fatty acids in lipids from blue-green algae

The main glycerolipids (monogalactosyl-, digalactosyl-, sulphoquinovosyl diacylglycerol, phosphatidylglycerol) from five blue-green algae (Microcystis, Anabaena, Nostoc, Oscillatoria, Tolypothrix) were analyzed for fatty acid composition, occurrence of diglyceride species and positional distribution of fatty acids between thesn-1- andsn-2-position of glycerol. In contrast to eucaryotic plants biosynthetically closely related lipids (monogalactosyl-, digalactosyl-, trigalactosyl diacylglycerol) show nearly identical diglyceride moieties, whereas sulphoquinovosyl diacylglycerol and phosphatidylglycerol are separated from galactolipids by composition as well as occurrence of fatty acids. On the other hand the positional distribution of fatty acids in all lipids is controlled exclusively by chain length and not by degree of unsaturation with C₁₈-fatty acids at thesn-1- and C₁₆-fatty acids at thesn-2-position. These results show that in procaryotic organisms the diversity in diglyceride portions of lipids is reduced as compared to eucaryotic organisms, but nevertheless does exist.Abbreviations MGD, DGD, TGD, SQD monogalactosyl-, digalactosyl-, trigalactosyl-, sulphoquinovosyl diacylglycerol - PG phosphatidyl glycerol     

Hymenobacter qilianensis sp. nov., isolated from a subsurface sandstone sediment in the permafrost region of Qilian Mountains,China and emended description of the genus Hymenobacter

A red–pink, Gram-negative, rod-shaped, non-motile, non-spore-forming bacterium, designated strain DK6-37 was isolated from the permafrost region of Qilian Mountains in northwest of China. Phylogenetic analysis based on 16S rRNA gene sequencing indicated that this isolate represents a novel member of the genus Hymenobacter, with low sequence similarities (<97 %) to recognized Hymenobacter species. Optimum growth was observed at 28 °C, pH 7.0 and 0 % NaCl. The strain was found to contain MK-7 as the predominant menaquinone. The polar lipids were identified as phosphatidylethanolanmine, two unknown aminophospholipids, one unknown aminolipid and three unknown polar lipids. The major fatty acids were identified as summed feature 3 (C_16:1 ω7c/C_16:1 ω6c as defined by MIDI), summed feature 4 (anteiso-C_17:1 B/iso-C_17:1 I), C_16:1 ω5c, iso-C_17:0 3-OH, iso-C_15:0 and C_18:0. The DNA G + C content was determined to be 67.4 mol %. On the basis of the polyphasic evidence presented, it is proposed that strain DK6-37 represents a novel species of the genus Hymenobacter, for which the name Hymenobacter qilianensis sp. nov. is proposed. The type strain is DK6-37^T (= CGMCC 1.12720^T = JCM 19763^T).     

Die Lipide von Endomycopsis vernalis bei verschiedener Stickstoff-Ernährung

1. Endomycopsis vernalis was cultivated on media with different N supply: series A 1%, series B 0,125% asparagine. Sonified cells were extracted and yielded 14.3% (A) and 65.3 (B) total lipids/non lipid dry matter respectively.
2. Neutral and complex lipids were separated by rubber membrane dialysis. There is no difference in the percentage of complex lipids of both series. The increase of lipids in cells grown on low N level is due to a higher content of neutral lipids.
3. Components of the neutral lipids, analysed by DC, were diglycerides, triglycerides, free and esterified ergosterol. Their percentage is influenced by the nutritional conditions. There is a significant increase of triglycerides and of sterol esters in the high lipid cells of series B.
4. Methyl esters of component fatty acids of glycerides and sterol esters were analyzed by GLC. Saturated acids C₁₄, C₁₅, C₁₆, C₁₇, C₁₈, monoenic acids C₁₆ and C₁₈, linoleic and linolenic acids were found to be present. Major acids were in all cases 18:1 (17–57%), 18:2 (18–50%) and 16:0 (10–18%). Linolenic acid is higher in di-and triglycerides of low lipid cells of series A than in high lipid cells of series B. Both qualitative and quantitative differences of fatty acids were found in sterol esters of series A and B respectively.
5. The major components of complex lipids, identified by DC and isolated by CC, in both series, were phosphatidyl choline (A:36.5, B:41.0%) and phosphatidyl ethanolamine (A:24.9, B:20.5%) in addition to small amounts of lysophosphatidyl choline, lysophosphatidyl ethanolamine, phosphatidyl serine, monophosphoinositide, diphosphatidyl glycerol and, possibly cerebroside like substances.
6. Methyl esters of the fatty acids of phosphatidyl choline and ethanolamine from both series were determined by GLC. In all samples 16:0, 18:0, 18:1, 18:2 and 18:3 acids were present besides of traces of 16:1 and 17:0. In contrast to neutral lipids the major acid of phospholipids is linoleic (53–58%), followed by oleic (8–24%) and linolenic acid (1–18%). The percentages of palmitic (4–8%) and stearic acids (tr.-1%) are small. Low lipid cells of series A differ from high lipid cells of series B by an increase of linolenic, and a decrease of linoleic acids, both in phosphatidyl choline and phosphatidyl ethanolamine.

     

Nonhongiella spirulinensis gen. nov., sp. nov., a bacterium isolated from a cultivation pond of Spirulina platensis in Sanya, China

A Gram-negative, aerobic, motile rod strain, designated Ma-20^T, was isolated from a pool of marine Spirulina platensis cultivation, Sanya, China, and was subjected to a polyphasic taxonomy study. Strain Ma-20^T can grow in the presence of 0.5–11 % (w/v) NaCl, 10–43 °C and pH 6–10, and grew optimally at 30 °C, pH 7.5–9.0 in natural seawater medium. The polar lipids were composed of phosphatidylethanolamine, three unidentified phospholipids and three unidentified polar lipids. The respiratory quinone was ubiquinone 8 (Q-8) and the major fatty acids were C_18:1ω6c/C_18:1ω7c (summed feature 8, 32.84 %), C_16:1ω6c/C_16:1ω7c (summed feature 3, 30.76 %), C_16:0 (13.54 %), C_12:03-OH (4.63 %), and C_12:0 (4.09 %). The DNA G+C content of strain Ma-20^T was 58 mol %. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain Ma-20^T belonging to Gammaproteobacteria, it shared 88.46–91.55 and 89.21–91.26 % 16S rRNA gene sequence similarity to the type strains in genus Hahella and Marinobacter, respectively. In addition to the large 16S rRNA gene sequence difference, Ma-20^T can also be distinguished from the reference type strains Hahella ganghwensis FR1050^T and Marinobacter hydrocarbonoclasticus sp. 17^T by several phenotypic characteristics and chemotaxonomic properties. On the basis of phenotypic, chemotaxonomic and phylogenetic properties, strain Ma-20^T is suggested to represent a novel species of a new genus in Gammaproteobacteria, for which the name Nonhongiella spirulinensis gen. nov., sp. nov. is proposed. The type strain is Ma-20^T (=KCTC 32221^T=LMG 27470^T).     

Differences in Lipid Composition between Undifferentiated and Mature Maize Chloroplasts

Lipid compositions of undifferentiated maize (Zea mays) chloroplasts, capable of fixing CO₂, were compared with the lipid compositions of mature chloroplasts, which do not fix CO₂, located in both the mesophyll and bundle sheath cells. The major lipids found in all three chloroplast types were the glycolipids, monogalactosyl diglyceride and digalactosyl diglyceride, followed by decreasing amounts of sulfolipid, phosphatidyl glycerol, phosphatidyl choline, phosphatidyl inositol, and diphosphatidyl glycerol. Quantitative differences in lipid components were observed among the chloroplast types. The mesophyll and bundle sheath maize chloroplasts differed in their chlorophyll a/chlorophyll b ratios (2.27 and 4.13 respectively) and their content of glycolipid relative to chlorophyll (51.8% glycolipid to 20.9% chlorophyll and 84.5% glycolipid to 10.1% chlorophyll respectively). A comparison between the lipid compositions of maize mesophyll chloroplasts and mesophyll chloroplasts obtained from spinach, sugar beet, and tobacco showed many similarities.     

Branched-chain amino acid metabolism controls membrane phospholipid structure in Staphylococcus aureus

Branched-chain amino acids (primarily isoleucine) are important regulators of virulence and are converted to precursor molecules used to initiate fatty acid synthesis in Staphylococcus aureus. Defining how bacteria control their membrane phospholipid composition is key to understanding their adaptation to different environments. Here, we used mass tracing experiments to show that extracellular isoleucine is preferentially metabolized by the branched-chain ketoacid dehydrogenase complex, in contrast to valine, which is not efficiently converted to isobutyryl-CoA. This selectivity creates a ratio of anteiso:iso C₅-CoAs that matches the anteiso:iso ratio in membrane phospholipids, indicating indiscriminate utilization of these precursors by the initiation condensing enzyme FabH. Lipidomics analysis showed that removal of isoleucine and leucine from the medium led to the replacement of phospholipid molecular species containing anteiso/iso 17- and 19-carbon fatty acids with 18- and 20-carbon straight-chain fatty acids. This compositional change is driven by an increase in the acetyl-CoA:C₅-CoA ratio, enhancing the utilization of acetyl-CoA by FabH. The acyl carrier protein (ACP) pool normally consists of odd carbon acyl-ACP intermediates, but when branched-chain amino acids are absent from the environment, there was a large increase in even carbon acyl-ACP pathway intermediates. The high substrate selectivity of PlsC ensures that, in the presence or the absence of extracellular Ile/Leu, the 2-position is occupied by a branched-chain 15-carbon fatty acid. These metabolomic measurements show how the metabolism of isoleucine and leucine, rather than the selectivity of FabH, control the structure of membrane phospholipids.     

Ultrastructure,composition of neutral lipids and their fatty acids of Candida tropicalis strain D-2 mutants resistant to the polyene antibiotic nystatin

This report deals with data on the cell ultrastructure of Candida tropicals strain D-2 mutants resistant to the polyene antibiotic, nystatin, and with an analysis of the fractional composition of neutral lipids and their fatty acids. The ultrastructural organization of the mutant cells is characterized by thickening of the cell wall and formation of invaginations into the cytoplasm, the appearance of new formations, large vacuoles, and reduction of the system of mitochondrial cristae. Lipids of nys^r mutants differ from those of the nys^s variant in having a decreased content of steroids and some fractions of neutral lipids. Certain nys^r mutants manifest difference in the relative amounts of saturated and unsaturated fatty acids (C_16:0, C_16:1, C_18:0, C_18:1).     

Photobacterium marinum sp. nov., a marine bacterium isolated from a sediment sample from Palk Bay,India

The novel, cream colored, Gram-staining-negative, rod-shaped, motile bacteria, designated strains AK15^T and AK18, were isolated from sediment samples collected from Palk Bay, India. Both strains were positive for arginine dihydrolase, lysine decarboxylase, oxidase, nitrate reduction and methyl red test. The major fatty acids were C_16:0, C_{18:1 ω7c}, C_{16:1 ω7c} and/or C_{16:1 ω6c} and/or iso-C_15:0 2-OH (summed feature 3). Polar lipids content of strains AK15^T and AK18 were found to bephosphatidylethanolamine (PE), two unidentified phospholipids (PL1 and PL2) and three unidentified lipids (L1-L3). The 16S rRNA gene sequence analysis indicated strains AK15^T and AK18 as the members of the genus Photobacterium and closely related to the type strain Photobacterium jeanii with pair-wise sequence similarity of 96.7%. DNA–DNA hybridization between strain AK15^T and AK18 showed a relatedness of 87%. Based on data from the current polyphasic study, strains AK15^T and AK18 are proposed as novel species of the genus Photobacterium, for which the name Photobacterium marinum sp. nov. is proposed. The type strain of Photobacterium marinum is AK15^T (=MTCC 11066^T = DSM 25368^T).