Cellular fatty acid composition ofCorallococcus coralloides

The fatty acid composition of five strains ofCorallococcus coralloides and three reference species ofMyxococcus were determined by gas-liquid chromatography. Methyl esters of fatty acid containing from 12 to 22 carbon atoms were identified. The major fatty acids present were C₁₅ and C₁₇ saturated branched chain, and both C₁₆ saturated and unsaturated straight chain acids. The C₁₇ saturated branched and straight chain acids, which were in valuable concentration in species ofMyxococcus, were not, however, detected in all strains ofC. coralloides. The application of these results in the distinction ofC. coralloides from the genusMyxococcus is discussed.     

FATTY ACIDS AND HYDROXY FATTY ACIDS IN THREE SPECIES OF FRESHWATER EUSTIGMATOPHYTES

The monocarboxylic fatty acids and hydroxy fatty acids of three species of freshwater microalgae—Vischeria punctata Vischer, Vischeria helvetica (Vischer et Pascher) Taylor, and Eustigmatos vischeri (Hulbert) Taylor, all from the class Eustigmatophyceae— were examined. Each species displayed a very similar distribution of fatty acids, the most abundant of which were 20:5n-3, 16:0, and 16:1n-7; C₁₈ polyunsaturated fatty acids were minor components. These fatty acid distributions closely resemble those found in marine eustigmatophytes but are quite distinct from those found in most other algal classes. These microalgae also contain long-chain saturated and unsaturated monohydroxy fatty acids. Two distinct types of hydroxy fatty acids were found: a series of saturated α-hydroxy acids ranging from C₂₄ to C₃₀ with a shorter series of monounsaturated α-hydroxy acids ranging from C₂₆ to C₃₀ together with a series of saturated β-hydroxy acids ranging from C₂₆ to C₃₀. The latter have not previously been reported in either marine or freshwater microalgae, although C₃₀ to C₃₄ midchain (ω-18)-hydroxy fatty acids have been identified in hydrolyzed extracts from marine eustigmatophytes of the genus Nannochloropsis, and C₂₂ to C₂₆ saturated and monounsaturated α-hydroxy fatty acids have been found in three marine chlorophytes. These findings have provided a more complete picture of the lipid distributions within this little studied group of microalgae as well as a range of unusual compounds that might prove useful chemotaxonomic markers. The functions of the hydroxy fatty acids are not known, but a link to the formation of the lipid precursors of highly aliphatic biopolymers is suggested.     

Fatty acids in green algae cultivated on a pilot-plant scale

Fatty acids fromChlorella vulgaris, Scenedesmus obliquus var.acutus and from a mixed culture of the two strains, Melnik, were converted to methyl esters, separated by gas chromatography, and identified by means of standards. The spectrum of fatty acids included both saturated and unsaturated acids (with odd and even numbers of carbon atoms) from C₁₂ to C₂₂. Fatty acids C_16:0, C_18:0 and C_20:3 were the major components in all cultures. Pure strains differed from the mixed culture in the production of C_18:1, C_12:0 and C_19:2 acids; the first of these was present in higher amounts in pure cultures only, the latter two being found in the mixed culture. The level of lipids was lower as compared to the literature data and their extractability was affected by the manner of preparation of algae and extraction conditions.     

Studies on the Changes of Meat Fats by Various Processings

Data are presented to show the gas chromatographic identification of a total of 18 saturated aliphatic γ- and δ-lactones obtained from melted beef depot fat, namely, δ-C₆, γ-C₇, γ-C₈, γ-C₉, and a homologous series of γ- and δ-lactones of the even-carbon numbers C₁₀ to C₁₆ and of smaller amount of the odd-carbon numbers C₁₁ to C₁₅. These lactones were isolated by steam distillation and silicic acid adsorption chromatography, and identified through gas chromatography and infrared spectroscopy.

Lactones obtained had a peach-like flavor, and it was suggested that lactones were important in heated beef fat as the flavor compounds.     

Distribution of C22-, C24- and C26-α-Unit-Containing Mycolic Acid Homologues in Mycobacteria

There are three mycolic acid homologues with C₂₂-, C₂₄- and C₂₆-α-units in Mycobacterium. In order to reveal the composition and distribution of these homologues in each subclass and molecular species of mycolic acids and to compare them with the composition of constitutive non-polar fatty acids (free and bound forms), we have separated non-polar fatty acids and each subclass of mycolic acids from 21 mycobacterial species by thin-layer chromatography, and analyzed non-polar fatty acid methyl esters by gas chromatography (GC) and the cleavage products of methyl mycolate by pyrolysis GC. We further performed mass chromatographic analysis of trimethylsilyl (TMS) ether derivatives of mycolic acid methyl esters by monitoring [B-29]⁺ ions (loss of CHO from the α-branched-chain structure of mycolic acids) of m/z 426, 454 and 482 which are attributed to C₂₂-, C₂₄- and C₂₆-α-units of TMS ether derivatives of methyl mycolates, respectively, (Kaneda, K. et al, J. Clin. Microbiol. 24: 1060-1070, 1986). By pyrolysis GC, C_22:0, C_24:0 and C_26:0 fatty acid methyl esters generated by the C₂-C₃ cleavage of C₂₂-, C₂₄- and C₂₆-α-unit-containing mycolic acid methyl esters, respectively, were detected. Their proportion was almost the same among subclasses of mycolic acids in every Mycobacterium and also similar to the proportion of constitutive non-polar C_22:0, C_24:0 and C_26:0 fatty acids. By mass chromatography, the composition and distribution of C₂₂- and C₂₄-α-unit-containing homologues were revealed to be similar between α- and α'-mycolic acids in every Mycobacterium. We further analyzed in detail M. vaccae and demonstrated that the mass chromatogram of C₂₂-α-unit-containing homologue was analogous in shape to that of the C₂₄-α-unit-containing one, with the latter mass chromatogram being up-shifted from the former by two carbon numbers, in every subclass of α-, α'-, keto and dicarboxy mycolic acids. The present study suggests that the compositions of three homologues of both mycolic acids and constitutive non-polar fatty acids, which are characteristic to each mycobacterial species, may reflect the proportion of the amount of free C_22:0, C_24:0 and C_26:0 fatty acids synthesized in the cell. It is further demonstrated that intermolecular condensation of two fatty acids which become α- and β-units of mycolic acids will occur independently of the carbon chain length or kinds of polar moieties of fatty acids.     

Changes in molecular species composition of nocardomycolic acids in nocardia rubra by the growth temperature

Nocardomycolic acids from Nocardia rubra were fully separated and characterized by a combination of argentation thin-layer chromatography and gas chromatography — mass spectrometry (GCMS). The occurrence of 20 or more different molecular species of mycolic acids was demonstrated. GCMS analysis of each subclass of mycolic acids after separation on AgNO₃ thin-layer chromatography revealed that in general the major species consisted of the even-carbon mycolic acids ranging from C₃₈ to C₅₂. However, the most abundant species differed by the subclasses; C₄₄ being in saturated, C₄₆ in monoenoic and C₄₆ in dienoic mycolic acids, respectively. All these acids were shown to possess C₁₂ or C₁₄ alkyl branch at 2 position, while double bonds were located in longer straight chain alkyl unit.By using this method, distinctive changes in mycolic acid composition by growth temperature were observed. The ratios of saturated, monoenoic to dienoic mycolic acids in a mixture of certain carbon numbered mycolic acids varied greatly, according to the shift of growth temperature. The mass fragmentographic analysis, monitoring M-15 ions derived from the loss of methyl group from the molecular ions showed the lower temperature (15°C) grown cells contained more unsaturated (especially dienoic) mycolic acids, while the higher temperature (40°C) grown cells contained more saturated mycolic acids in both extractable and cell-wall bound lipids. These changes in mycolic acid composition occurred shortly after shifting up the growth temperature from 20°C to 43°C at a logarithmic stage of the bacterial growth.     

Elongation of mitochondrial fatty acids during brain development

Abstract— Elongation of mitochondrial fatty acids was studied in whole brain samples from rats before, during and after the period of myelination. The mitochondria were isolated by centrifugation in a discontinuous sucrose gradient and incubated under N₂ in a medium containing NADH, NADPH, ATP and acetyl-[1-¹⁴C]coenzyme A. Fatty acids were extracted, methylated and analysed by gas-liquid chromatography. A distinct pattern emerged in which brain mitochondria from rats undergoing myelination synthesized longer chain fatty acids preferentially, particularly C_22:4. Mitochondria from brains of mature rats synthesized shorter chain fatty acids preferentially, mainly C_18:0 and C_20:4. We suggest that eicosamonoenoic acid (C_22:1) is a precursor in vivo of nervonic acid (C_24:1).     

Determination of structure and composition of suberin from the roots of carrot, parsnip, rutabaga, turnip, red beet, and sweet potato by combined gas-liquid chromatography and mass spectrometry

Suberin from the roots of carrots (Daucus carota), parsnip (Pastinaca sativa), rutabaga (Brassica napobrassica), turnip (Brassica rapa), red beet (Beta vulgaris), and sweet potato (Ipomoea batatas) was isolated by a combination of chemical and enzymatic techniques. Finely powdered suberin was depolymerized with 14% BF₃ in methanol, and soluble monomers (20-50% of suberin) were fractionated into phenolic (<10%) and aliphatic (13-35%) fractions. The aliphatic fractions consisted mainly of ω-hydroxyacids (29-43%), dicarboxylic acids (16-27%), fatty acids (4-18%), and fatty alcohols (3-6%). Each fraction was subjected to combined gas-liquid chromatography and mass spectrometry. Among the fatty acids very long chain acids (>C₂₀) were the dominant components in all six plants. In the alcohol fraction C₁₈, C₂₀, C₂₂, and C₂₄ saturated primary alcohols were the major components. C₁₆ and C₁₈ dicarboxylic acids were the major dicarboxylic acids of the suberin of all six plants and in all cases octadec-9-ene-1, 18-dioic acid was the major component except in rutabaga where hexadecane-1, 16-dioic acid was the major dicarboxylic acid. The composition of the ω-hydroxyacid fraction was quite similar to that of the dicarboxylic acids; 18-hydroxy-octadec-9-enoic acid was the major component in all plants except rutabaga, where equal quantities of 16-hydroxyhexadecanoic acid and 18-hydroxyoctadec-9-enoic acid (42% each) were found. Compounds which would be derived from 18-hydroxyoctadec-9-enoic acid and octadec-9-ene-1, 18-dioic acid by epoxidation, and epoxidation followed by hydration of the epoxide, were also detected in most of the suberin samples. The monomer composition of the six plants showed general similarities but quite clear taxonomic differences.     

Ultrastructure and lipid chemistry of specialized epidermal structure of Indian porcupines and hedgehog

Poddar‐Sarkar, M., Raha, P., Bhar, R., Chakraborty, A. and Brahmachary, R.L. 2011. Ultrastructure and lipid chemistry of specialized epidermal structure of Indian porcupines and hedgehog. —Acta Zoologica (Stockholm) 92 : 134–140. In the present study, we investigated the ultrastructural variations of specialized epidermal structure of Indian porcupines (Hystrix indica and Atherurus macrourus) and hedgehog (Hemiechinus collaris) as well as the variation in the fatty acid composition of total lipid fraction. Scanning electron microscope images reveal the usual scaly structure in surface view and network of channels in cross‐section but with different orientation of partition walls. The lipid profile reveals the presence of free sterol, long‐chain alcohol, free fatty acids, wax ester and sterol ester in all the three cases and trace amount of triglyceride, diglyceride and monoglyceride. Gas chromatography–mass spectrometry analysis of fatty acid methyl ester of total lipid fraction indicates the presence of C₈‐C₂₂ fatty acids in Hystrix indica, C₈‐C₁₈ in Atherurus macrourus and C₈‐C₂₀ fatty acids in Hemiechinus collaris. It is interesting to note that the total lipid fraction of hedgehog shows no branched‐chain, unsaturated and odd‐carbon fatty acids. Odd‐carbon fatty acid and branched‐chain fatty acids detected in the adult H. indica but were absent in juvenile H. indica as well as in A. macrourus. With the exception of C_18:1, the other unsaturated fatty acids were also absent in both juvenile H. indica and A. macrourus.     

Determination of C20-C30 fatty acids by reversed-phase chromatographic techniques: An efficient method to quantitate minor fatty acids in serum of patients with adrenoleukodystrophy

An analytical method for the determination of saturated very long chain (VLC) fatty acids in the serum has been devised. Free fatty acids obtained after hydrolysis of total lipid extracts were converted intop-bromophenacyl esters. The derivatives were purified in two sequential steps by clean-up on C₁₈ reversed-phase cartridge and fractionation by reversed-phase thin-layer chromatography (TLC), and then quantitated by high performance liquid chromatography (HPLC) analysis. This technique provides a reliable and alternative method for the biochemical identification of patients and carriers of an inherited metabolic disease characterized by the accumulation of saturated VLC fatty acids (C₂₄–C₂₆) such as Adrenoleukodystrophy (ALD). In four cases of diagnosed ALD the fatty acid composition of serum total lipids was dramatically enriched in saturated VLC fatty acids compared to controls. The ratio of hexacosanoic acid (C₂₆₀) to docosanoic acid (C₂₂₀) in ALD patients was approximately six-fold higher than that of healthy controls or patients affected by metabolic or neurological disorders other than ALD.     

Untersuchungen an Beauveria tenella (NRRL 2334, 2335, 2336; bisher Agaricus campestris)

Zusammenfassung Die physikalischen und chemischen Kennzahlen und Daten des durch Ätherextraktion gewonnenen Rohlipids von Beauveria tenella wurden bestimmt und spektrale Absorptionskurven aufgenommen.Die Methylester der Fettsäuren wurden gaschromatographisch analysiert. Die einzelnen Fettsäuren wurden identifiziert und ihr mengenmäßiger Anteil ermittelt.Etwa 90% der Fettsäuren haben eine Kettenlänge von 16–18 Kohlenstoffatomen. Nahezu 60% der Fettsäuren sind ungesättigt; der prozentuale Anteil dieser Säuren nimmt mit steigender Zahl der Doppelbindungen ab.Octadecensäure hatte mit 25,6% den größten Anteil an der Gesamtmenge.An seltenen Fettsäuren konnte unter anderem eine gesättigte C₁₇- und C₂₄-Fettsäure nachgewiesen und das Vorkommen einer gesättigten C₁₉-und C₂₁-Fettsäure wahrscheinlich gemacht werden.

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Summary The physical and chemical constants and the spectral absorption curves of crude lipids of Beauveria tenella, obtained by means of etherextraction, were determined.The methyl esters of the fatty acids were analyzed using gas-liquid chromatography. The individual fatty acids were identified and the amount of each in the complete sample determined.Approximately 90% of the fatty acids had a 16–18 carbon-chain length, while almost 60% of the fatty acids were unsaturated. The percentage of fatty acids with unsaturated bonds was reciprocally proportional to the number of double bonds present.Octadecenoic acid comprised 25,6% of the total fatty acids and represented the largest single amount of a specific fatty acid present.With respect to unusual fatty acids, a saturated C₁₇ and a C₂₄ fatty acid were identified, while the presence of a C₁₉ and a C₂₁ fatty acid was indicated.

     

Isoflavone,wax and triterpene constituents of Wyethia mollis

The hexane extract of Wyethia mollis contains the n-alkanes C₁₅-C₁₈, C₂₀-C₂₅, C₂₇ and C₂₉. Linoleic acid was the only detectable acidic component. A mass spectral analysis of the wax ester fraction indicated that it was a mixture of homologues, the saturated even-carbon acids n-C₁₆-C₃₀ esterfield with the saturated even-carbon alcohols n-C₁₈-C₂₆. The chloroform extract yielded the known isoflavones santal and 3′-O-methylorobol along with a new lanostane-type triterpene, 22,25-epoxy-lanosta-7:9(11)-dien-3-one. The wide distribution of n-alkanes and the decreased odd-even carbon ratio are consistent with the proposed primitive nature of this plant.     

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.     

Comparative studies on simple lipids ofSclerotium cepivorum,the causal agent of white rot of onion,and other fungi of the class Deuteromycetes

Summary The simple lipids ofSclerotium cepivorum, the causal agent of white rot of onion and nine other fungal species of the same class were investigated. The fatty acid composition of the simple lipids of these fungi were determined by GLC. The main fatty acids common to these fungal species were C₁₆ (saturated) and C₁₈ (unsaturated) acids. The sterol fraction was isolated by column chromatography and its components were detected by GLC and mass spectrometry. Ergosterol and γ-Ergostenol were found mostly in all fungal species under investigation. However, two fungal species namelyAlternaria alternata andScolecobasidium constrictum showed no Ergosterol.     

Differences in cellular fatty acids and lipid composition inClostridium pasteurianum under nitrogen-and non-nitrogen-fixing conditions

Clostridium pasteurianum total cellular saturated fatty acids increased through its growth cycle from 81% to 91% but varied significantly in the composition under nitrogen- and non-nitrogen-fixing conditions. During ammonia-assimilating growth, palmitic acid decreased from 67.7% to 43.5% by late log while marked increases in shorter chain saturated fatty acids (C_15:0 and below) and a long chain saturated C_22:0 occured. In contrast, under N₂-fixing growth conditions, palmitic acid increased from 45.5% to 84.3% by late log, representing nearly the total amound of saturated fatty acids found inC. pasteurianum. The total cellular lipid concentration decreased as the culture aged. irrespective of the nitrogen sources; however, the phospholipid concentration increased significantly during N₂-fixing growth as compared with a 50% decrease during ammonia-assimilating conditions. The implication of these differences and possible role of palmitic acid and phospholipids inC. pasteurianum nitrogen fixation process are discussed.     

The effects of 2,4-dinitrophenol on the oxidation of fatty acids byPseudomonas aeruginosa

Summary The effects of varying concentrations of 2,4-dinitrophenol on the oxidation of a series of saturated, aliphatic fatty acids byPs. aeruginosa were studied. The oxidation of C₄, C₆, C₈, C₁₀, C₁₂, C₁₃, C₁₄ and C₁₆ acids is stimulated by certain concentrations of this inhibitor. However, the concentration of 2,4-dinitrophenol which causes stimulation of oxygen uptake with capric acid does not produce an increase in numbers of the organisms in a medium containing the fatty acid as the sole carbon source. This investigation was supported by a Fellowship from the Anderson Oil and Chemical Company, Inc., Portland, Connecticut.     

Identification by gas chromatography and mass spectrometry of lipids from the rat Harderian gland

Lipids from rat Harderian glands were extracted with ethyl acetate, hydrolysed with base and examined by gas chromatography (GC) and gas chromatography—mass spectrometry (GC—MS) as trimethylsilyl (TMS), [²H₉]TMS, methyl ester—TMS, picolinyl, nicotinate and nicotinylidene derivatives. The latter three derivatives were used to reveal the structures of the alkyl chains of fatty acids, alcohols and glycerol ethers, respectively. Forty-eight compounds were identified, representing about 97% of the total extracted lipids as measured by GC peak areas. The major constituents were fatty acids with chain lengths from 12 to 22 carbon atoms (mainly C₁₈ and C₂₀) and fatty alcohols (C₁₆ to C₂₆) derived from wax esters. Most of these acids and alcohols were unsaturated in the ω-7 position and were accompanied by smaller amounts of the saturated and ω-5 monounsaturated analogues. Glycerol ethers were also identified for the first time in this secretion; the ether chains contained from 14 to 19 carbon atoms (mainly 16) and were straight-chain saturated, unsaturated (ω-5 and ω-7) and branched (iso). The only sterol found was cholesterol amounting to 1.24% of the total extract.     

The distribution of fatty acids including petroselinic and tariric acids in the fruit and seed oils of the Pittosporaceae, Araliaceae, Umbelliferae, Simarubaceae and Rutaceae

The fatty acid composition of the fruit oils or seed oils of Pittosporaceae (eight genera, 10 species), Araliaceae (two species), Simarubaceae (three species), and of one umbelliferous and one rutaceous species were determined by gas chromatography, argentation TLC and ozonolysis. In the Pittosporaceae, in which the major C₁₈ fatty acid of all species was either oleic acid (18:1, 9c) or linoleic acid (18:2, 9c, 12c), large amounts of C₂₀ and C₂₂ fatty acids seem to occur regularly. Petroselinic (18:1, 6c) and tariric (18:1, 6a) acids were absent. However, petroselinic acid was the major fatty acid in the Araliaceae and Umbelliferae. In these two families only small amounts of C₂₀ and C₂₂ acids were detected and tariric acid was absent. The Rutales contained relatively high amounts of trans-octadecenoic acids (18:1, 9t). Tariric acid was the major fatty acid in the two species of Picramnia (Simarubraceae), which also contained small amounts of petroselinic acid. The major fatty acids in Ailanthus glandulosa (Simarubaceae) and Phellodendron amurense (Rutaceae) were linoleic or linolenic acid (18:3, 9c, 12c, 15c); these species contained neither tariric nor petroselinic acid and the levels of C₂₀ and C₂₂ fatty acids were low. The appearance of schizogenous resin canals and polyacetylenes and the absence of iridoids and petroselinic acid allows the Pittosporaceae to be separated from the Rutales and Araliales and to be placed in an independent order, the Pittosporales. Arguments for a rather close relationship of the Pittosporales to the Araliales and Cornales (including the Escalloniaceae) are presented.     

Content of fatty acids ofChlorella kessleri in a deep tank fermentation

Chlorella kessleri cultivated in a deep tank contained 4.8% of non-polar lipid; 51% of this fraction represents saturated fatty acids, 7% unsaturated fatty acids. Our investigation of the fatty acids profile demonstrated even- and odd-numbered saturated and unsaturated fatty acids ranging from C₁₂ to C₂₀. Unlike in otherChlorella species, stearic acid was the dominant fatty acid found. Also shown was an elevated C_16:0 fatty acid content and a reduced level of unsaturated fatty acids.     

Esters of trehalose from Corynebacterium diphtheriae: A modified purification procedure and studies on the structure of their constituent hydroxylated fatty acids

The purification procedure of 6,6′-diesters of trehalose from Corynebacterium diphtheriae was modified and the isolated substance was analysed by mass spectrometry as its permethylated derivative. The fatty acid moiety released from the glycolipid after alkaline hydrolysis was studied by mass spectral analysis of the O-methylated and O-acetylated methyl ester derivatives. By argentation thin-layer chromatography, three species of O-acetylated methyl esters were recognized, corresponding to saturated, mono-unsaturated and di-unsaturated α-branched-β-hydroxylated fatty acids. The double bond was located by ozonolysis of the O-acetylated methyl ester derivatives, by gas chromatography of the reaction product and mass spectrometry of the effluent from the gas chromatograph. The main components of each species of α-branched-β-hydroxylated fatty acids found in the gly colipid fraction of C. diphtheriae were 2-tetradecyl-3-hydroxyoctadecanoic acid (C₃₂H₆₄O₃, corynomycolic acid), 2-tetradecyl-3-hydroxy-11-octadecenoic acid (C₃₂H₆₂O₃, corynomycolenic acid), 2-tetradec-7′-enyl-3-hydroxy octadecanoic acid (C₃₂H₆₂O₃) and 2-tetradec-7′-enyl-3-hydroxy-11-octadecenoic acid (C₃₂H₆₀O₃, corynomycoldienic acid). The glycolipid fraction from C. diphtheriae is obviously a complex mixture of 6,6′-diesters of trehalose.