On the chemical structure of the apical droplet from eggs of Culex pipiens

The chemical nature of the apical droplet from eggs of Culex pipiens was investigated by chromatographic techniques. Results indicated that the hydrolysate of the apical drop contains C-12, C-14, C-16, and C-18 straightchain aliphatic fatty acids. A C-12β-hydroxy fatty acid was also found, but the largest component of the fatty acid mixture of the apical drop was shown to be a C-14β-OH fatty acid. Two other fractions appear to be unsaturated fatty acids, probably C-12 and C-14. Quantitative estimation of the percentage of each fatty acid in the mixture showed that about 85 per cent of the fatty acid content of the apical drop consisted of hydroxy fatty acids. By thin-layer chromatography, the largest component coincided with β-OH myristic acid.Glycerol was confirmed to be present in the hydrolysate. Feeding studies with radioactive ³²PO₄^?3 and ³⁵SO₄^?2 showed no significant incorporation of phosphorus, but a sulphur-containing anionic compound could be detected in the apical drop. Infrared analysis showed the presence of an ester group, double bond, primary and secondary alcohol groups, suggesting the presence of hydroxy-, unsaturated-, saturated straight-chain fatty acids, as well as mono-and diglycerides. The structural evidence explains in part the surfactant properties of the apical drop.     

Identification of the Basic Structure of a Glycolipid from Selenomonas ruminantium as β-Glucosaminyl-1,6-glucosamine

Lipid materials extracted by chloroform-methanol from solvent and acid treated cells of Selenomonas ruminantium grown with ¹⁴C-valerate were analyzed by thin-layer chromatography. At least 12 radioactive lipid materials were present in the extract. The major compound, designated as spot A, carried approximately 70% of radioactivity of this fraction. The spot A compound was purified by column and thin-layer chromatography and its chemical structure was studied. The basic structural unit of this material was tentatively identified as β-gIucosaminyl-1,6-glucosamine with O-acyl and amide linked fatty acids.     

Synthesis of β-hydroxy fatty acids and β-amino fatty acids by the strains of Bacillus subtilis producing iturinic antibiotics

The iturinic antibiotics, which contain long chain β-amino acids, are produced by Bacillus subtilis. Screening these strains for the presence of a possible precursor of the iturinic antibiotics, we isolated a lipopeptide containing β-hydroxy fatty acids. The structure of this compound was studied and it appears to be identical or structurally very similar to surfactin. The carbon chain of its β-hydroxy fatty acids was n C₁₆, iso C₁₆, iso C₁₅ or anteiso C₁₅. The percentages of each β-hydroxy fatty acids varied according to the strain producing iturinic antibiotics and were influenced by addition of branched-chain α-amino acids to the culture medium. These results demonstrate for the first time that iso C₁₄ β-hydroxy fatty acid is a constituent present in such a surfactin like lipopeptide. Besides, the presence of radioactive β-hydroxy fatty acids in the phospholipids when the strains were grown in the presence of sodium [¹⁴C]acetate seems also characterize the different strains producing iturinic antibiotics.     

Anaerobic Transformation of Alkanes to Fatty Acids by a Sulfate-Reducing Bacterium, Strain Hxd3

Strain Hxd3, an alkane-degrading sulfate reducer previously isolated and described by Aeckersberg et al. (F. Aeckersberg, F. Bak, and F. Widdel, Arch. Microbiol. 156:5-14, 1991), was studied for its alkane degradation mechanism by using deuterium and ¹³C-labeled compounds. Deuterated fatty acids with even numbers of C atoms (C-even) and ¹³C-labeled fatty acids with odd numbers of C atoms (C-odd) were recovered from cultures of Hxd3 grown on perdeuterated pentadecane and [1,2-¹³C₂]hexadecane, respectively, underscoring evidence that C-odd alkanes are transformed to C-even fatty acids and vice versa. When Hxd3 was grown on unlabeled hexadecane in the presence of [¹³C]bicarbonate, the resulting 15:0 fatty acid, which was one carbon shorter than the alkane, incorporated a ¹³C label to form its carboxyl group. The same results were observed when tetradecane, pentadecane, and perdeuterated pentadecane were used as the substrates. These observations indicate that the initial attack of alkanes includes both carboxylation with inorganic bicarbonate and the removal of two carbon atoms from the alkane chain terminus, resulting in a fatty acid one carbon shorter than the original alkane. The removal of two terminal carbon atoms is further evidenced by the observation that the [1,2-¹³C₂]hexadecane-derived fatty acids contained either two ¹³C labels located exclusively at their acyl chain termini or none at all. Furthermore, when perdeuterated pentadecane was used as the substrate, the 14:0 and 16:0 fatty acids formed both carried the same numbers of deuterium labels, while the latter was not deuterated at its carboxyl end. These observations provide further evidence that the 14:0 fatty acid was initially formed from perdeuterated pentadecane, while the 16:0 fatty acid was produced after chain elongation of the former fatty acid with nondeuterated carbon atoms. We propose that strain Hxd3 anaerobically transforms an alkane to a fatty acid through a mechanism which includes subterminal carboxylation at the C-3 position of the alkane and elimination of the two adjacent terminal carbon atoms.     

Fatty-Acid Composition of Candida utilis as Affected by Growth Temperature and Dissolved-Oxygen Tension

Analyses were made of the fatty-acid composition of Candida utilis NCYC 321 grown in a chemostat at a dilution rate (equal to growth rate) of 0.1 hr⁻¹ and at temperatures in the range of 30 to 15 C and dissolved oxygen tensions between 75 and <1 mm of Hg. Cells grown under glucose limitation or NH₄⁺ limitation contained mainly C_16:0, C_16:1, C_18:0, C_18:1, C_18:2, and C_18:3 acids as detected by gas-liquid chromatography of methyl esters of the acids from lipids extracted with chloroform-methanol. The relative proportions of these acids varied with the growth temperature and the dissolved-oxygen tension in the culture. A decrease in growth temperature from 30 to 20 C led to an increased synthesis of unsaturated acids in cells grown under either limitation at a fixed-oxygen tension in the range of 75 to 5 mm of Hg. In cultures with a dissolved-oxygen tension of 1 and <1 mm of Hg, a further decrease in temperature to 15 C caused an increased synthesis of unsaturated fatty acids. A decrease in dissolved-oxygen tension led to a diminished synthesis of unsaturated fatty acids in cells grown at a fixed temperature under either limitation. Cells grown at a fixed temperature under glucose limitation synthesized a greater proportion of C₁₆ acids at the expense of C₁₈ acids as the dissolved oxygen tension was decreased from 75 to <1 mm of Hg. A preferential synthesis of C₁₆ acids also occurred as the growth temperature was decreased from 30 to 15 C in cells grown under glucose limitation at a fixed-oxygen tension. The same effect was observed in cells grown under NH₄⁺ limitation when the temperature was lowered from 30 to 20 C; but when the temperature was decreased further to 15 C, the cells synthesized a slightly greater proportion of C₁₈ acids. Synthesis of a large proportion of C₁₆ acids was accompanied by an excretion of pyruvate, and occasionally traces of 2-ketoglutarate, and an increased intracellular accumulation of certain amino acids.     

In vitro synthesis of mycolic acids by the fluffy layer fraction of Bacterionema matruchotii

Biosynthetic activity for mycolic acid occurred in the fluffy layer fraction but not in the 5000g supernatant of Bacterionema matruchotii. With [1-¹⁴C]palmitic acid as precursor for the in vitro system, the predominant product was identified as C_32:0 mycolic acid by radio-gas-liquid chromatographie (radio-GLC) and gas chromatographic/mass spectroscopic analyses; if [1-¹⁴C]stearic acid was used, two major radioactive peaks appeared on GLC: one corresponding to the peak of (C_34:0 + C_34:1) mycolic acids and the other to (C_36:0 + C_36:1) mycolic acids. By pyrolysis/radio-GLC analysis, C_32:0 mycolic acid synthesized by [1-¹⁴C]palmitic acid was pyrolyzed at 300 °C to form palmitaldehyde (the mero moiety) and methyl palmitate (the branch moiety). The pH optimum for the incorporation of [1-¹⁴C]palmitate into bacterionema mycolic acids was 6.4 and the reaction required a divalent cation. The in vitro system utilized myristic, palmitic, stearic and oleic acids (probably via their activated forms) well as precursors, among which myristic and palmitic acids were more effective than the rest. Avidin showed no effect on the biosynthesis of mycolic acid from ¹⁴C-palmitate whereas cerulenin, a specific inhibitor of β-ketoacyl synthetase in de novo fatty acid synthesis, inhibited the reaction at a relatively higher concentration. Thin-layer chromatographic analysis of lipids extracted from the reacting mixture without alkaline hydrolysis showed that both exogenous [1-¹⁴] fatty acid and synthesized mycolic acids were bound to an unknown compound by an alkali-labile linkage and this association seemed to occur prior to the condensation of two molecules of fatty acid.     

Initial Reactions in Anaerobic Alkane Degradation by a Sulfate Reducer, Strain AK-01

An alkane-degrading, sulfate-reducing bacterial strain, AK-01, isolated from a petroleum-contaminated sediment was studied to elucidate its mechanism of alkane metabolism. Total cellular fatty acids of AK-01 were predominantly C even when it was grown on C-even alkanes and were predominantly C odd when grown on C-odd alkanes, suggesting that the bacterium anaerobically oxidizes alkanes to fatty acids. Among these fatty acids, some 2-, 4-, and 6-methylated fatty acids were specifically found only when AK-01 was grown on alkanes, and their chain lengths always correlated with those of the alkanes. When [1,2-¹³C₂]hexadecane or perdeuterated pentadecane was used as the growth substrate, ¹³C-labeled 2-Me-16:0, 4-Me-18:0, and 6-Me-20:0 fatty acids or deuterated 2-Me-15:0, 4-Me-17:0, and 6-Me-19:0 fatty acids were recovered, respectively, confirming that these monomethylated fatty acids were alkane derived. Examination of the ¹³C-labeled 2-, 4-, and 6-methylated fatty acids by mass spectrometry showed that each of them contained two ¹³C atoms, located at the methyl group and the adjacent carbon, thus indicating that the methyl group was the original terminal carbon of the [1,2-¹³C₂]hexadecane. For perdeuterated pentadecane, the presence of three deuterium atoms, on the methyl group and its adjacent carbon, in each of the deuterated 2-, 4-, and 6-methylated fatty acids further supported the hypothesis that the methyl group was the terminal carbon of the alkane. Thus, exogenous carbon appears to be initially added to an alkane subterminally at the C-2 position such that the original terminal carbon of the alkane becomes a methyl group on the subsequently formed fatty acid. The carbon addition reaction, however, does not appear to be a direct carboxylation of inorganic bicarbonate. A pathway for anaerobic metabolism of alkanes by strain AK-01 is proposed.     

Ratios of Carbon Isotopes in Microbial Lipids as an Indicator of Substrate Usage

The occurrence and abundance of microbial fatty acids have been used for the identification of microorganisms in microbial communities. However, these fatty acids can also be used as indicators of substrate usage. For this, a systematic investigation of the discrimination of the stable carbon isotopes by different microorganisms is necessary. We grew 11 strains representing major bacterial and fungal species with four different isotopically defined carbon sources and determined the isotope ratios of fatty acids of different lipid fractions. A comparison of the differences of δ¹³C values of palmitic acid (C_16:0) with the δ¹³C values of the substrates revealed that the isotope ratio is independent of the growth stage and that most microorganisms showed enrichment of C_16:0 with ¹³C when growing on glycerol. With the exception of Burkholderia gladioli, all microorganism showed depletion of ¹³C in C_16:0 while incorporating the carbons of glucose, and most of them were enriched with ¹³C from mannose, with the exception of Pseudomonas fluorescens and the Zygomycotina. Usually, the glycolipid fractions are depleted in ¹³C compared to the phospholipid fractions. The δ¹³C pattern was not uniform within the different fatty acids of a given microbial species. Generally, tetradecanoic acid (C_14:0) was depleted of ¹³C compared to palmitic acid (C_16:0) while octadecanoic acid (C_18:0) was enriched. These results are important for the calibration of a new method in which δ¹³C values of fatty acids from the environment delineate the use of bacterial substrates in an ecosystem.     

Die Fettsäuren in ganzen Zellen,Thylakoiden und Lipopolysacchariden von Rhodospirillum rubrum und Rhodopseudomonas capsulata

Zusammenfassung Die photosynthetischen Bakterien Rhodospirillum rubrum und Rhodopseudomonas capsulata wurden auf ihre Fettsäurezusammensetzung untersucht. Die Hauptfettsäuren von R. rubrum waren C_16:0 (11%), C_16:1 (30%) und C_18:1 (52%). Vaccensäure (C_18:1) bildete 94% der Fettsäuren von Rps. capsulata. Anaerobe Lichtzellen (thylakoidhaltig) unterschieden sich nicht in ihrem Fettsäuremuster von aeroben Dunkelzellen (thylakoidfrei). Gereinigte Thylakoide aus Lichtzellen zeigten das gleiche Fettsäuremuster wie die ganzen Zellen.Nach Phenol/Wasser-Extraktion der ganzen Zellen bei 68° C war bei beiden Organismen sowohl aus Licht- als auch aus Dunkelzellen eine Substanz aus der gäßrigen Phase isolierbar, welche in den Sedimentationseigenschaften mit den Lipopolysacchariden der Enterobacteriaceae übereinstimmte und nach orientierenden Untersuchungen Zucker enthält. Aus ihr wurde ein Fettsäuregemisch gewonnen, dessen Zusammensetzung von dem aus ganzen Zellen erheblich abwich. In Rps. capsulata enthielt es C_12:1 (40%) und C_16:0 (50%), während in R. rubrum sich das Fettsäuremuster über den Bereich von C₁₀ bis C₂₀ erstreckte. Licht- und Dunkelzellen wiesen in dieser Substanz Unterschiede in der Fettsäurezusammensetzung auf. Der quantitative Anteil der Fettsäuren in dieser Substanz, bezogen auf die Gesamtfettsäuren der Zelle, betrug in Licht- und Dunkelzellen 5–7%. Hydroxy-myristinsäure ließ sich in beiden Organismen nicht nachweisen.

---

Fatty acid composition of whole cells, thylakoids and lipopolysaccharides of Rhodospirillum rubrum and Rhodopseudomonas capsulata

Summary The fatty acid composition of the photosynthetic bacteria Rhodospirillum rubrum and Rhodopseudomonas capsulata was investigated. The bulk of fatty acids of R. rubrum consisted of C_16:0 (11%), C_16:1 (30%), and C_18:1 (52%). The major fatty acid of Rps. capsulata was vaccenic acid (C_18:1), which accounted for 94% of the total fatty acids. Cells of both organisms, which were grown anaerobically in the light and fitted out with thylakoids had the same fatty acid composition as cells grown aerobically in the dark, which have no thylakoids.Purified thylakoids had the same fatty acid pattern as whole cells. Whole cells of light and dark cultures were extracted with phenol/water at 68° C. An opalizing fraction in the aqueous phase was sedimentable in the ultracentrifuge like the lipopolysaccharides of the Enterobacteriaceae. The pattern of fatty acids in this compound differed considerably from that of whole cells. The major fatty acids in this macromolecular fraction were C_12:1 (40%) and C_16:0 (50%) in Rps. capsulata, whereas in R. rubrum the whole range of fatty acids from C₁₀ to C₂₀ was demonstrable. Light and dark grown cells differed in the fatty acid composition of that compound. The fatty acid content of the extracted fraction accounted for 5–7% of the total fatty acids of whole cells. No hydroxymyristic acid could be identified in either R. rubrum or Rps. capsulata.

     

IN VIVO METABOLISM OF [G-3H]LIGNOCERIC ACID IN DEVELOPING RAT BRAIN1

Abstract— [G-³H]Lignoceric acid (tetracosanoic acid) was injected into the brains of 20-day-old rats, and the animals were killed after 8, 24, or 72 h. Various lipids were isolated from these brains, and the distribution of radioactivity was determined. The injected free acid rapidly disappeared, and the radioactivity was incorporated into varying chain-length nonhydroxy- and hydroxy saturated fatty acids of sphingolipids and phospholipids. Little radioactivity was found in unsaturated acids, sphingo-sine, and cholesterol. A time-dependent shift of the label among various fatty acids was relatively small 8 h after injection, probably because of the metabolic stability of the brain sphingolipids. In cerebrosides, the radioactivity was equally distributed between nonhydroxy and x-hydroxy fatty acids of all chain lengths. C₂₃ and C₂₂ fatty acids contained equal total radioactivities; C₂₃ and C₂₄ fatty acids contained similar specific activities. These results confirm the significant role of a-hydroxylation and 2-oxidation in the synthesis of very long-chain fatty acids in brain. In total lipid fatty acids, docosanoic acid (22:0) contained more radioactivity than its α-oxidation precursor, α-hydroxytricosanoic acid (23h:0) at all times. In sphingolipid fatty acids, the specific activity of 21:0 was always higher than that of its ct-oxidation precursor 22:0. These observations indicate that part of the 22:0 and 21:0 was derived by β-oxidation from the injected lignoceric acid or its α-oxidation product, respectively.     

Transformation of sterols byMycobacterium vaccae: effect of lecithin on the permeability of cell envelopes to sterols

An enhancement of Β-sitosterol transformation to androstendione byMycobacterium vaccae observed in medium containing egg-yolk lecithin, was associated with the incorporation of a considerable amount of lecithin into the cell envelope lipids. By GC/MS measurements, fatty acids ranging from 14 to 22 carbon atoms were identified in the lipids removed from the cells by organic solvents. Octadecenoic (18:1), 2-methyl-octadecenoic (2-Me 18:1), and hexadecanoic (16:0) acids were the major components of the lipid preparation obtained from both the control cells, and the cells grown in lecithin-containing medium. However, in the fatty acid pattern of the latter a distinct increase in the C_18:1 component, concomitant with the decrease in the 2-Me 18:1 fatty acid was demonstrated. The C₁₆ fatty acid fraction also showed a higher content of methyl-branched components in the control cell preparation. The enrichment in unsaturated fatty acids increases fluidity of lipids, whereas the decrease in methyl-branched fatty acids may affect the conformation of the surface lipidic components, which may result in enhanced sterol penetration through the cell wall barrier in the presence of lecithin.     

Biodegradation and conversion of alkanes and crude oil by a marine Rhodococcus

A hydrocarbon degrader isolated from a chronically oil-polluted marine site was identified as Rhodococcus sp. on the basis of morphology, fatty acid methyl ester pattern, cell wall analysis, biochemical tests and G + C content of DNA. It degraded upto 50% of the aliphatic fraction of Assam crude oil, in seawater supplemented with 35 mM nitrogen as urea and 0.1 mM phosphorus as dipotassium hydrogen orthophosphate, after 72 h at 30 ° and 150 revolutions per minute. The relative percentage of intracellular fatty acid was higher in hydrocarbon-grown cells compared to fructose-grown cells. The fatty acids C₁₆ , C16_{16 :1} C₁₈ and C_{18 : 1} were constitutively present regardless of the growth substrate. In addition to these constitutive acids, other intracellular fatty acids varied in correlation to the hydrocarbon chain length supplied as a substrate. When grown on odd carbon number alkanes, the isolate released only monocarboxylic acids into the growth medium. On even carbon number alkanes only dicarboxylic acids were produced.     

Production of fatty acids and lipid by a Candida sp. growing on a fraction of n-alkanes predominating in tridecane

A Candida sp. was grown on a fraction of n-alkanes (dodecane 22%, tridecane 48%, tetradecane 28%) as sole carbon source. The growth rate was increased most markedly by using high concentrations of n-alkanes (16.7% v/v). When grown in a 5 liter fermentor, the yeast reached its highest yield (60 g. of cell dry wt/l) with a concomitant high yield of fatty acids (21 g of fatty acids/l), by using a nitrogen-deficient medium. To achieve good growth, it was essential to use an inoculum (1 part into 10) of rapidly growing cells and beneficial to increase the agitation rate gradually once growth had begun. After 108 hr maximum conversions of substrate to product were: 71.5% (w/w) for alkanes into cells and 24.8% (w/w) for alkanes into fatty acids. Of the, total fatty acids at the end of the fat-accumulating phase of growth 54% were shorter in chain length than palmitic acid (C₁₆H₃₂O₂). When grown on glucose, as sole carbon source, less than 2% of the total fatty acids were shorter than palmitic acid. When n-alkanes were added to cells growing on glucose, short-chain fatty acids (C₁₀ to C₁₄) were synthesized immediately, indicating a derepressed enzyme system for hydrocarbon assimilation and the absence of diauxie. The production of these acids was at the apparent sacrifice of linoleic acid synthesis. In spite of the high conversion ratios, it is concluded that it would be uneconomical to produce fatty acids, even expensive ones such as lauric acid, by microbial transformation of n-alkanes.     

Bacterial Degradation of N-Lauroyl-L-valine

Studies were conducted on the degradation of N-lauroyl-L-valine by type cultured bacteria. Many strains could utilize sodium N-lauroyl-L-valinate as carbon and nitrogen sources for their growth. Metabolism of N-lauroyl-L-valine was investigated in detail using Ps. aeruginosa AJ2116. Laurie acid was identified by gas chromatography suggesting cleavage of N-acyl linkage in N-lauroyl-L-valine.

Laurie acid might be metabolized to capric acid (C₁₀) and caprylic acid (C₈) becuase the accumulated substances gave nearly identical peaks with those of authentic fatty acids on gas chromatograms. The experiment using N-lauroyl-L-valine (¹⁴C) indicated that ¹⁴CO₂ was produced as a final product. Valine was not detected because it might be metabolized very rapidly immediately after its release.

It was supposed that the enzymes or enzyme systems degrading N-lauroyl-L-valine might be constitutive from the experiment using two kinds of cells grown in the medium containing N-lauroyl-L-valine or nutrient broth.     

Photosynthesis,lipids and proteins in the cyanobacteriumSynechocystis PCC 6803 as affected by temperature

The cyanobacteriumSynechocystis PCC 6803 was grown photoautotrophically in an inorganic medium at constant growth temperatures of 20, 38 (control) or 43°C for 9 h. The up and down-shift of cultivation temperature decreased the growth as measured by culture absorbance and chlorophylla content. However, high temperature slightly increased the oxygen evolution while temperature lower than control inhibited oxygen evolution during the whole incubation period. The protein synthesis studied by¹⁴C-labeled protein declined under low temperature by about 50%. The fatty acid pattern is characterized as lacking in C₂₀/C₂₂ acids but containing large amounts of C₁₆ and C₁₈ polyunsaturated fatty acids, 16:2 and 18:3 in particular. The lower temperature increased the percentage of monounsaturated fatty acids while higher temperature increased the saturated fatty acid content in total lipids and lipid classes studied.     

Variation in fatty acid composition of <Emphasis Type="BoldItalic">Arthrospira</Emphasis> (Spirulina) strains

A study of the fatty acid composition was made for 35 Arthrospira strains, concentrating on the most abundant fatty acids, the two polyunsaturated C₁₈ acids, linoleic and γ-linolenic acid, and palmitic acid. When grown at 30 ^∘C and low irradiance (10 μmol photon m⁻² s⁻¹), these three acids together formed 88–92% of total fatty acids. There were considerable differences in the composition of the two polyunsaturated acids. Depending on the strain, linoleic acid formed 13.1–31.5% and γ-linolenic acid formed 12.9–29.4% total fatty acids. In contrast, the range for palmitic acid was narrow: 42.3–47.6% of total fatty acids. Repeat experiments on several strains under defined conditions led to closely similar results for any particular environment, suggesting that fatty acid composition can be used as an aid in differentiating between strains. Five additional strains, which had apparently originated from the same original stock cultures as 3 of the 35 in the main study, but from different culture collections, were also assayed. With four strains the results were similar, irrespective of culture source, but with one strain marked differences occurred, especially in the polyunsaturated C₁₈ fatty acid fraction. These differences were independent of the age of the culture. In addition, straight morphotypes derived during repeat subcultures of four strains; each showed a similar fatty acid composition to that of the helical morphotypes of the same strains. A decrease in temperature from 30 to 20 ^∘C, an increase in irradiance (at 30 ^∘C) from 10 to 70 μmol photon m⁻² s⁻¹ and transfer to dark heterotrophy all favoured an increase in polyunsaturated C₁₈ fatty acids. The highest γ-linolenic acid content of any conditions was found for three strains grown heterotrophically on glucose in the dark at 30 ^∘C. A comparative study of six strains of Spirulina confirmed a previous study showing the absence of γ-linolenic acid in all Spirulina strains, thus permitting the separation of these two genera.     

Fatty acid composition of lipids from differentiated tissues and cell cultures of Euonymus europaeus

The fatty acid patterns of Euonymus europaeus callus cultures and cell suspension cultures were analysed at the beginning of stationary growth phase and compared with those from the respective differentiated tissues. The lipid and fatty acid patterns in cell cultures differed remarkably from those in the tissues of the mother plant. No glycerol triacetate was detected in the callus cultures derived from differentiated tissues whereas in seeds this lipid compound amounts to 29%. In addition to fatty acids normally occurring in differentiated tissues, lipids in cultured cells also contained short-chain (C₁₂–C₁₄) as well as very long-chain fatty acids (C₂₀–C₂₄). In tissue culture cells the major fatty acids were found to be saturated, whereas in the mother cells unsaturated fatty acids were predominant. Palmitic acid is the most abundant fatty acid in most of the cultures. Lauric, myristic and palmitic acid amount to 50% in lipids of cell suspension cultures.     

Characterization of the Glycolipid Associated with Alzheimer Paired Helical Filaments

Abstract: In the present study, analytical techniques including gas chromatography/mass spectrometry (GC/MS)-assisted carbohydrate linkage-analysis, one- and two-dimensional NMR, and matrix-assisted laser desorption/ionization time of flight mass spectroscopy (MALDI-MS) have been used to characterize the structure of the glycolipid associated with the paired helical filaments (PHF) isolated from the neurofibrillary tangles of Alzheimer's diseased brain. The ¹H NMR spectrum of acid-hydrolyzed protein-resistant core PHF (prcPHF) displays resonances that can be assigned to fatty acid and glucose. There are no resonances present that would indicate the presence of protein, amino acids, or a sphingosine base. Using two-dimensional homonuclear correlated spectroscopy, homonuclear Hartmann-Hahn, and heteronuclear multiple quantum coherence experiments, resonances in the ¹H and ¹³C NMR spectrum of native PHF were assigned to a nonreducing terminal α-1,6-glycosidically linked glucose, an internal α-1,6-linked glucose, and an α-1,2,6-linked glucose. The narrow line-widths observed for these residues suggest that they arise from glucose residues undergoing rapid segmental motion. The carbohydrate portion of the PHF-associated glycolipid was analyzed using GC/MS linkage analysis and confirmed the presence of terminal and internal α-1,6-linked glucose and α-1,2,6-linked glucose in a molar ratio of 2:1:1. Three components of the PHF-associated glycolipid fraction having masses 2,416, 2,325, and 2,237 Da were observed using MALDI-MS. The least abundant, heavier mass component (2,416 Da) was best fit to a structure with a tridecamer of glucose having a single esterified C₂₀ fatty acid (Glc₁₃ + C₂₀ or Glc₁₃ + C_20:1), whereas the more abundant, lower mass components were best fit to noncovalently associated glycolipid dimers, each with a glucose pentamer or hexamer having two C₁₄, C₁₆, or C₁₈ esterified fatty acids {D[(Glc₅ + C₁₈) + (Glc₆ + C₁₆)] or D[(Glc₅ + C₁₄) + (Glc₆ + C₁₄)]}. The ratio of glucose to fatty acid calculated from these best-fit structures of the more abundant mass components (5.5 ± 1.1:1.0) is in reasonable agreement with the same ratio calculated from peak integrations in the NMR spectra of acid-hydrolyzed prcPHF (6.2 ± 1.6). Structural similarities between PHF-associated glycolipid and other glycolipid amphiphiles known to form PHF-like filaments indirectly suggest that this unique glycolipid may be an integral component of the PHF suprastructure.     

Purification and characterization of mononocardomycoloylglycerol from Nocardia rhodochrous

A component of the acetone-soluble lipids of Nocordia rhodochrous grown on glycerol, was purified by column chromatography on silicic acid and characterized by infrared, nuclear magnetic resonance spectroscopy, optical rotation measurement and product identification after alkaline hydrolysis. Glycerol was the sole water-soluble component and nocardomycolic acids with chain lengths ranging from C₄₀ to C₄₄ were the constituent fatty acids identified. On the basis of the evidence obtained, the substance isolated from N. rhodochrous is identified as a mixture of mononocardomycoloylglycerols in which nocardomycolic acids are bound to one of the primary hydroxyl groups of the glycerol molecule.     

Fatty acid exchange of phospholipids in membranes of rat heart

Incorporation of ¹⁴C fatty acids in phospholipids of plasma membranes and sarcoplasmic reticulum of rat heart was studied. Mainly phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were labelled. Our studies showed that the incorporation of unsaturated fatty acids (oleic and linoleic acid) was higher than for saturated fatty acids (palmitic and stearic acid). The range of uptake was between 0.2 and 2.2 nmol·mg⁻¹ protein·h⁻¹ and 0.5–7.4 nmol·μgatom⁻¹ P₁·h⁻¹, respectively. Uptake of activity in individual phospholipids (measured after separation on TLC) was calculated as percentage of total activity. Incorporation in phosphatidylcholine was higher than in phosphatidylethanolamine. Phosphatidylcholine showed an increasing sequence for the following fatty acids: C_18:0 < C_16:0 < C_18:0 < C_18:2. However, phosphatidylethanolamine showed a decreasing sequence for the incorporation of the same fatty acids. Labelling of PC was always greater than for PE, except for stearic acid which was better incorporated into phosphatidylethanolamine. Uptake of the same fatty acid into phospholipids of sarcoplasmic reticulum was always higher than uptake into plasma membranes. As incorporation of fatty acids bound to albumin was studied in isolated membranes of rat heart, the addition of ATP and CoASH was an absolute requirement.