Untersuchungen zur Anreicherung von Ubichinon-9 aus Lipid-Kohlenwasserstoff-Franktionen mittels Sepahdex LH-20

Sephadex LH-20 was tested for a potential application in the enrichment of ubiquinone-9 from a lipid-hydrocarbon-extract. The lipid-hydrocarbon-extract was isolated from yeast grown on gas oil (b. p. 240 to 380°C) as carbon-source. The main components of the lipid-hydrocarbon extract were phosphatides, glycerides, fatty acids, and hydrocarbons of gas oil. By use of Sephadex LH-20 and CHCl₃/CH₃OH (2:1)as eluent it is possible to concentrate the whole ubiquinone-9 in a special fraction both directly from the lipid-hydrocarbon-extract and from the aceton-soluble part of the extract The concentration of ubiquinone in the special fraction was ten times higher than in the raw-material and the hydrocarbons were separated completely. Ubiquinone-9 can be obtained by a simplified separation from the enriched fraction.     

Untersuchungen zur Anreicherung von Ubichinon-9 aus Lipid-Kohlenwasserstoff-Fraktionen mittels Kurzwegdestillation

Short path distillation was used for the enrichment of ubiquinone-9 from a lipid hydrocarbon extract. The main components of the lipid hydrocarbon extract were hydrocarbons of gas oil (b. p. 513 to 653 K), phosphatides, glycerides, and fatty acids. Phosphatides were isolated by extraction with acetone before distillation. The acetone soluble fraction was distilled by a pressure of ≦ 0.5 kPa and a temperature of 433 to 453 K. The concentration of the ubiquinone-9 in the residue of distillation was three times higher than in the acetone soluble fraction. Ubiquinone-9 can be obtained by a simplified separation from the enriched fraction.     

Polyprenols of Aspergillus niger. Their characterization, biosynthesis and subcellular distribution

A polyprenol complex of Aspergillus niger was shown, by using spectrometric methods, to consist of a family of exo-methylene-hexahydroprenols that contain between 18 and 24 isoprene residues per molecule. Each prenol contains two trans residues, three saturated residues (alpha, omega and psi) and an exo-methylene substituent on the carbon atom beta to the isopropyl group in each omega-residue. The ubiquinone complex consisted of 90% ubiquinone-9, 9% ubiquinone-8 and 1% ubiquinone-10. The amount of polyprenol complex present reached a maximum of 1.7mg/culture bottle after 9-10 days of growth, coincident with the maximum weight of mycelium. The amount of ergosterol (10mg/culture bottle) and ubiquinone (1mg/culture bottle) reached a peak at 8 days. By the 13th day of growth the yield of ergosterol had fallen by 20% and that of ubiquinone by 85%. A study of the incorporation of [2-(14)C]mevalonate over different time-intervals confirmed that there was a slow turnover of prenol, a more rapid turnover of ergosterol and a very rapid turnover of ubiquinone. At any one time each member of the prenol complex had essentially the same specific radioactivity as other members of the complex. A similar conclusion was made about the ubiquinone mixture. Just over half of the polyprenol present was esterified to fatty acids. Subcellular fractionation studies indicated that the unesterified prenol is associated primarily with a mitochondrial fraction, whereas the ester is more widely distributed.     

Wearing a raincoat: exocrine secretions contain anti-wetting agents in the oribatid mite, <Emphasis Type="Italic">Liacarus subterraneus</Emphasis> (Acari: Oribatida)

Liacarus subterraneus is a large, soil-dwelling oribatid mite species that possesses a conspicuously shiny, clean and not wettable cuticular surface. The exocrine cuticular chemistry of this species was investigated by means of gas chromatography–mass spectrometry. Besides a fraction of hydrocarbons and a terpene, hexane extracts of whole mite bodies exhibited free carboxylic acids and their glycerides as main components. The compounds were arranged in three distinct extract profiles. Based on data from individual extracts, (1) the majority (more than 3/4) of specimens showed large amounts of 1,2-dioctanoyl-glycerol (and three other related esters) but no (or only traces of) free carboxylic acids. (2) In about 1/8 of extracts, free acids (mainly octanoic (caprylic) acid) and glycerides were detected. This second type of profile highly varied with respect to the relative abundance of acids and esters. (3) The third profile (in about 7% of specimens) exclusively exhibited free acids and no (or only traces of) glycerides. In addition, a few extracts exhibited no components at all. The extract compounds most likely originate from the lipid layer of the cerotegument of L. subterraneus. The cuticle of individuals that possessed extractable cerotegumental compounds (profile I, II, III) exhibited strong water repellent properties, while the cuticle of individuals that possessed no components in their extract did not. After hexane extraction, water repellent properties got lost. The distinct extract profiles detected most likely portray the stepwise generation of an anti-wetting, exocrine surface lipid layer of glycerides: If this layer is lost, fatty acids may be discharged again (profile III) and may subsequently esterify (profile II) to larger and more stable esters (diacyl-glycerols), eventually building up the “raincoat” (mainly profile I) of L. subterraneus.     

Untersuchungen zur chemischen Zusammensetzung der Lipidfraktion von Acinetobacter calcoaceticus

Lipids were extracted from the cells of Acinetobacter calcoaceticus EB 10 C IMET B 395 grown on gas oil (Bp. 240–360 °C) with benzine/alcohol (80 : 20). The lipid-hydrocarbon-extract obtained by this extraction method was 18.4%. The extract was composed of hydrocarbons, waxes, phospholipids, fatty acids, glycerides, and ubiquinones. The main components among the lipids were waxes. The compositions of phospholipid, fatty acid, wax, and ubiquinone fractions were analysed.     

THE BIOGENESIS OF MITOCHONDRIA : III. The Lipid Composition of Aerobically and Anaerobically Grown Saccharomyces cerevisiae as Related to the Membrane Systems of the Cells

The growth conditions known to influence the occurrence of mitochondrial profiles and other cell membrane systems in anaerobic cells of S. cerevisiae have been examined, and the effect of the several growth media on the lipid composition of the organism has been determined. The anaerobic cell type containing neither detectable mitochondrial profiles nor the large cell vacuole may be obtained by the culture of the organism on growth-limiting levels of the lipids, ergosterol, and unsaturated fatty acids. Under these conditions, the organism has a high content of short-chain saturated fatty acids (10:0, 12:0), phosphatidyl choline, and squalene, compared with aerobically grown cells, and it is especially low in phosphatidyl ethanolamine and the glycerol phosphatides (phosphatidyl glycerol + cardiolipin). The high levels of unsaturated fatty acids normally found in the phospholipids of the aerobic cells are largely replaced by the short-chain saturated acids, even though the phospholipid fraction contains virtually all of the small amounts of unsaturated fatty acid present in the anaerobic cells. Such anaerobic cells may contain as little as 0.12 mg of ergosterol per g dry weight of cells while the aerobic cells contain about 6 mg of ergosterol per g dry weight. Anaerobic cell types containing mitochondrial profiles can be obtained by the culture of the organism in the presence of excess quantities of ergosterol and unsaturated fatty acids. Such cells have increased levels of total phospholipid, ergosterol, and unsaturated fatty acids, although these compounds do not reach the levels found in aerobic cells. The level of ergosterol in anaerobic cells is markedly influenced by the nature of the carbohydrate in the medium; those cells grown on galactose media supplemented with ergosterol and unsaturated fatty acids have well defined mitochondrial profiles and an ergosterol content (2 mg per g dry weight of cells) three times that of equivalent glucose-grown cells which have poorly defined organelle profiles. Anaerobic cells which are low in ergosterol synthesize increased amounts of squalene.     

Measurement of Hydroxy and Hydroperoxy Fatty Acids by a High Pressure Liquid Chromatography with a Column-switching System

Hydroxy and hydroperoxy fatty acids were labeled with 9-bromomethylacridine at room temperature. They were separated from the degradation products and less polar fatty acid derivatives on an octyl silicagel column, and put on an octadecyl silicagel column by on-line column switching. By this method, picomolar levels of the derivatives were measured with good reproducibility. The detection limit of 16-hydroxy-hexadecanoic acid as a representative was 0.9 pmol (S/N =3) and the relative standard deviation of its peak areas was 2.5% (18.5 pmol, n = 7). The method was used for the measurement of hydroxy fatty acids derived from hydroperoxy fatty acids and phosphatidylcholine (PC) hydroperoxides spiked in human plasma. By incubation at 37°C for 4h with human plasma, the hydroperoxy fatty acid was reduced to the corresponding hydroxy fatty acid. In this condition, the PC hydroperoxides showed a considerable decrease, however, a small portion (2.5–3%) of PC hydroperoxides decomposed gave the corresponding hydroxy fatty acids.     

The effects of some amino acids on growth and lipid production in Inonotus obliquus grown in vitro

The effect of the distribution of free and esterified fatty acids, including triterpenes and sterols, on the mycelial growth of the basidiomycete Inonotus obliquus (Pers. ex Fr.) Pilat in malt extract and solid supplemented mineral media was investigated. The amino acids DL-alanine. DL-aspartic acid, L-glutamic acid, L-leucine, L-methionine, and L-serine were added to the cultures at nitrogen concentrations of 0.1, 0.5 and 0.9 g/l, respectively. Differences in the average mycelial growth were distinct at concentrations of 0.5 g/l and 0.9 g/l. L-Methionine had the highest inhibitory effect on growth. At all concentrations, the amino acids reduced the production of lanosterol. The other identified triterpenes were inotodiol, 3β,hydroxy-lanosta-8,24-dien-21-al, 3β,21-dihydroxy-lanosta-8,24-diene, trametenolic acid, and ergosterol peroxide. The main fatty acids were either C 16:0 or C 18:2 both free or in glycerides.     

Untersuchungen zur chemischen Zusammensetzung der Lipidfraktion von Lodderomyces elongisporus EH 15

Lipids were extracted from the cells of Lodderomyces elongisporus EH 15 grown on gas oil (Bp. 240 to 360 °C) with benzine/alcohol (80:20). The lipid-hydrocarbon-fraction obtained by this extraction method was 18.5%. It was composed of hydrocarbons, phospholipids, fatty acids, glycerides, sterols, ubiquinones, and vitamines. The main components among the lipids were phospholipids. The compositions of phospholipid, fatty acid, sterol, and ubiquinone fractions were analysed.     

Studies on the Browning of Kori-tofu

Properties of ether extract of browned Kori-tofu were investigated. More than half amount of neutral glycerides of Kori-tofu changed into weak-acidic substances after browning, in which most of extracted brown pigments were contained. These weak-acidic substances showed properties of carbonyl and reductone, and mainly contained glycerides, whose constitutional fatty acids were palmitic, oleic, and linoleic acids as well as a little amount of azelaic and stearic acids.

From the ether extract of browned Kori-tofu were isolated and identified azelaic acid and carbonyls. The carbonyls identified were benzaldehyde, methylethylketone, crotonal, acetaldehyde, α-ketoaldehydes of C₂, C₃, C₄, C₅, C₇ and C₉, and hexane-2,3-dione. Two nitrogeneous carbonyl compounds were isolated. Formation mechanisms of these compounds and their roles in browning of Kori-tofu were discussed.     

Reductive and oxidative biosynthesis of plasmalogens in myelinating brain

Palmitic acid-1-(14)C and hexadecanol-1-(14)C were administered intracerebrally to 18-day-old rats. Incorporation of radioactivity into the constituent alkyl, alk-1-enyl, and 1-acyl moieties, as well as into the 2-acyl moieties, of the ethanolamine phosphatides of brain was determined after 1, 2, 3, 6, and 22 hr. Incorporation of radioactivity from hexadecanol into both alkyl ethers and alk-1-enyl ethers proceeded at a rate more than 10 times higher than from palmitic acid. Hexadecanol was rapidly oxidized to fatty acids which were incorporated into the acyl moieties of the ethanolamine phosphatides. When palmitic acid was used as a precursor, labeled long-chain alcohols could be isolated from the lipid extract. As labeled long-chain aldehydes could not be detected in any of the lipid extracts, alcohols appear to be key intermediates for the biosynthesis of both alkyl and alk-1-enyl glycerophosphatides.     

High-temperature gas chromatography-mass spectrometry for skin surface lipids profiling

Skin surface lipids (SSLs) arising from both sebaceous glands and skin removal form a complex lipid mixture composed of free fatty acids and neutral lipids. High-temperature gas chromatography coupled with electron impact or chemical ionization mass spectrometry was used to achieve a simple analytical protocol, without prior separation in classes and without prior cleavage of lipid molecules, in order to obtain simultaneously i) a qualitative characterization of the individual SSLs and ii) a quantitative evaluation of lipid classes. The method was first optimized with SSLs collected from the forehead of a volunteer. More than 200 compounds were identified in the same run. These compounds have been classified in five lipid classes: free fatty acids, hydrocarbons, waxes, sterols, and glycerides. The advantage to this method was it provided structural information on intact compounds, which is new for cholesteryl esters and glycerides, and to obtain detailed fingerprints of the major SSLs. These fingerprints were used to compare the SSL compositions from different body areas. The squalene/cholesterol ratio was used to determine the balance between sebaceous secretion and skin removal. This method could be of general interest in fields where complex lipid mixtures are involved.     

TOTAL LIPID PRODUCTION OF THE GREEN ALGA NANNOCHLOROPSIS SP. QII UNDER DIFFERENT NITROGEN REGIMES1

The green alga Nannochloropsis sp. QII was cultivated in media with sufficient and growth-limiting levels of nitrogen (nitrate). Nitrogen deficiency promoted lipid synthesis yielding cells with lipids comprising 55% of the biomass. The major lipids were triacylglycerols (79%), polar lipids (9%) and hydrocarbons (2.5%). The polar lipids consisted of a broad range of phospholipids, glycolipids and sulfolipids. Other lipids identified were pigments, free fatty acids, saponifiable and unsaponifiable sterol derivatives, various glycerides, a family of alkyl-1, 4-dioxane derivatives and a series of alkyl- and hydroxyalkyl-dimethyl-acetals. Experiments in which ¹⁴CO₂ was provided at different times in the growth cycle demonstrated that enhanced lipid biosynthesis at low nitrogen levels resulted principally from de novo CO₂ fixation.     

Lipid Composition of Soil Amoebae

SYNOPSIS. Lipid content of axenic cultures of 3 species of soil amoebae was investigated. The strains studied were Acanthamoeba sp., Hartmannella rhysodes and Mayorella palestinensis. No appreciable differences were apparent in the amount of sterols and free fatty acids present in the different strains. The sterols were "fast acting" and were present mostly in free form. There was a difference in the quantities of glycerides, Acanthamoeba sp. containing the highest amount. Sterol of H. rhysodes was isolated and identified as ergosterol by chemical and physical criteria. The implications of these findings are discussed.     

Distribution of newly formed fatty acids among glycerolipids of isolated perfused rat liver.

Livers of chow fed rats were perfused 1-3 h with buffer, glucose, albumin, and red blood cells, made up in 100 percent D(2)O. Glycerolipids were isolated and the deuterated fatty acids determined by gas chromatography with mass spectrometry on Silar 5 CP. Percentage of replacement by deuterated acids ranged from 1 to 14, of which palmitate was 87 percent. Differences were found in total lipid class and in subcellular distribution of the newly synthesized acids. Microsomes had 37 percent more deuterated acids than the total or floating fat. At 3 h the highest replacement was found in diacylglycerols (17 percent) and free fatty acids (11 percent). Of the palmitate in hepatic choline and ethanolamine phosphatides, 6.9 percent and 4.7 percent, respectively, contained dueterium. The serine and inositol phosphatides had a higher proportion of deuterated palmitate (7.7 percent) than other phosphatides. The data support the hypothesis that palmitate is incorporated into glycerolipids largely via de novo synthesis while stearate enters them by deacylation-acyl transfer replacement.     

Constituents of the basidiomycete Scleroderma aurantium

The following compounds were isolated from the extract of the fresh peridium of Scleroderma aurantium: palmitic acid, linoleic acid, a mixture of triglycerides, an ester of a steroidal diol, ergosterol, ergosterol peroxide, 9(11)-dehydroergosterol peroxide, lanosta-8,23- dien-3β,25-diol, lanosta-8,24-diene-3β,23-diol, and mannitol.     

Neutral Lipids and Phospholipids of Free-Living and Bacteroid Forms of Two Strains of Rhizobium Infective on Lotus pedunculatus

The neutral lipids and phospholipids of two strains of rhizobia in their free-living state and in symbiosis with a host plant are described. The principal lipid classes found were the polymer poly-beta-hydroxybutyrate, phospholipids, free fatty acids, glycerides, methyl esters, aliphatic alcohols, and hydrocarbons. The lipids include unusual unsaturated methyl-branched and saturated methoxy-branched fatty acids. Most components were found to be common to both forms of both strains, although the proportions varied. A number of strain differences could be discerned.     

Ubiquinone-10 Protects Neurons from Virus-Induced Degeneration

Abstract: Cultured neurons from rat dorsal root ganglia and cerebral cortex were infected with Sendai virus, which gives a productive replication with lysis of most neurons, and with the RW strain of mumps virus, which undergoes defective replication causing degeneration of only 30–40% of the neurons within 5 days after initial infection. In Sendai virus-infected cells the amount of polyisoprenoid lipids was enhanced. In mumps virus-infected cultures there were transient reductions in the contents of cholesterol, dolichol, and ubiquinone-9 in the cultures, whereas the reduction in the ubiquinone-10 level was progressive, reaching 20% of its original value 21 days after infection. Treatment of mumps virus-infected cultures with ubiquinone-10 protected the neurons from degeneration, whereas no effects were observed on exposure to ubiquinone-9. Linolenic acid (18:3) and arachidonic acid (20:4), but not myristic acid (14:0) and palmitic acid (16:0), also had significant neuroprotective effects.     

Distribution of newly formed palmitate and stearate among molecular species of choline and ethanolamine phosphatides.

The choline and ethanolamine phosphatides derived from isolated rat livers during perfusion with 75 percent deuterated water (Kuksis, A., Myher, J.J., Marai L., Yeung, S.K.F., Steiman, I. & Mookerjea, S. (1975) Can. J. Biochem. 53, 509-518) were resolved into molecular species by argentation thin-layer chromatography. The time course of percentage replacement of the newly synthesized fatty acids in each molecular species was determined by gas chromatography with mass spectrometry. The results confirmed the earlier postulated differential utilization of palmitic and stearic acids in glycerolipid biosynthesis as well as supported the hypothesis of a precursor-product relationship between the oligoenoic and tetranoic species of both phosphatides. Calculations of half-lives gave values of 14-19 h for palmitoyl oligoenes, 40-50 h for palmitoyl tetraenes, and 22-28 h for palmitoyl hexaenes of both choline and ethanolamine phosphatides. The corresponding stearoyl species had half-lives which ranged from 89 to 200 h. Evidence was obtained for a metabolic heterogeneity among subsets of molecular species recognized on the basis of combinations of new and old glycerol and fatty acids in the same glycerolipid molecule.     

Enzynatic method of disrupting yeast cell walls]

The influence of the preparation of mucolytic enzymes obtained from the culture liquid filtrate of Actinomyces griseinus on the yeast Candida guilliermondii was investigated. Lipids, hydrocarbons and proteins were extracted and free amino acids were separated from the biomass made of disrupted yeast cells. The extract contained 73% of cytoplasmic substances whereas normal yeast (enzymically untreated) contain on the average 13.7%. Lipids and hydrocarbons were measured.