Fatty acid composition and the classification of the Porifera

Investigation of the fatty acid profiles of 55 species of Porifera has confirmed the occurrence of high percentages of both long chain fatty acids (C₂₄-C₃₀) and polyunsaturated acids. These features of the fatty acid profile in conjunction with the content of branched chain acids and the dominance of particular acids in different species allow some systematic discussion. The Dictyoceratida, Clathriidae, Halichondrida, Homoscleromorpha and Calcarea are distinguished by aspects of their fatty acid profiles. The diversity in number and type of sponge fatty acids exceeds that of any other phylum. Environmentally induced variation in fatty acid content is such that percentage compositions alone have little taxonomic informational value.     

Fatty acid composition of sterol esters from Citrus sinensis,C. paradisi,C. limon aurantifolia and C. limettioides sacs

The fatty acid compositions of sterol esters from 4 citrus species, viz, orange. grapefruit, lemon and lime, were determined by GLC. Each species possessed its own intrinsic fatty acid pattern which could be used to differentiate it from the other species. In most cases varieties within a species had fatty acid patterns which could be used for varietal differentiation. In all citrus tested except Columbia lime, the major acid was linoleic acid; this acid varied from 10 to 56% of the total acid content. The ratios of 16/16:1 were distinct for each citrus species. The C₂₂-C₂₉ fatty acids were prevalent in citrus sterol esters ranging from 6·5% for some orange and grapefruit varieties to over 41% for two lime varieties. In all varieties C₂₄ was the most prominent of these longer chain fatty acids. Argentation TLC indicated that these longer chain fatty acids primarily were esterified to dimethyl sterols. ft*|One of the laboratories of the Southern Region, Agricultural Research Service, U.S. Department of Agriculture.     

Changes in fatty acid composition of lipid classes in developing mustard seed

Maturation of mustard (Sinapis alba) seed proceeds with a sharp decrease in the amounts of palmitic and linoleic acids in the total lipids up to 6 weeks after flowering (WAF). Concomitantly, the concentration of oleic acid increases, reaching a plateau at 4 WAF, which is followed by chain elongation of oleic acid to gadoleic and erucic acids. Compositional changes in constituent fatty acids of individual lipid classes indicate that the very long-chain monounsaturated fatty acids (C₂₀ and C₂₂), as opposed to common long-chain fatty acids (C₁₆ and C₁₈), are metabolized to triacylglycerols mainly by esterification to preformed diacylglycerols and monoacylglycerols, rather than via esterification to glycerol-3-phosphate or lysophosphatidic acids.     

Composition of the epicuticular and intracuticular wax layers on Kalanchoe daigremontiana (Hamet et Perr. de la Bathie) leaves

Epicuticular and intracuticular waxes from both adaxial and abaxial surfaces of the leaves of Kalanchoe daigremontiana were analyzed. All wax mixtures were found to contain approximately equal amounts of triterpenoids and very long chain fatty acid (VLCFA) derivatives. The triterpenoid fraction consisted of glutinol (8-19% of the total wax) and friedelin (4-9%), together with smaller amounts of glutanol, glutinol acetate, epifriedelanol, germanicol and β-amyrin. The VLCFA derivatives comprised C₂₇-C₃₅ alkanes (19-37% of the total wax), C₃₂-C₃₄ aldehydes (3-7%), C₃₂ and C₃₄ fatty acids (0.2-3%), C₂₆-C₃₆ primary alcohols (4-8%), and C₄₂-C₅₂ alkyl esters (2-9%). The wax layers were found to differ in triterpenoid amounts, with the intracuticular wax containing higher percentages of most triterpenoids than the epicuticular wax. Friedelin, the only triterpenoid ketone present, showed the opposite distribution with higher proportions in the epicuticular wax. VLCFA derivatives also accumulated to higher percentages in the epicuticular than in the intracuticular wax layer. Epicuticular wax crystals were observed on both the adaxial and abaxial leaf surfaces.     

Elongation of acyl-CoAs by microsomes from etiolated leek seedlings

Long chain fatty acid synthesis was studied using etiolated leek seedling microsomes. In the presence of ATP, [2-¹⁴C]malonyl-CoA was incorporated into fatty acids of C₁₆C₂₆. The omission of ATP, even in the presence of acetyl-CoA, led to a complete loss of activity, which was restored by addition of exogeneous acyl-CoAs. Comparison of acyl-CoA (C₁₂C₂₄) elongation showed that stearoyl-CoA, in the presence of [2-¹⁴C]malonyl-CoA, was the more efficient precursor leading to the formation of fatty acids having a chain length of C₂₀C₂₆. [1-¹⁴C]C₁₆CoA and [1-¹⁴C]C₁₈CoA were elongated in the presence of malonyl-CoA, without degradation of the acyl chain. The time-course and the malonyl-CoA concentration curves showed that [1-¹⁴C]C₁₈CoA was a better primer than [1-¹⁴C]C₁₆CoA. Acyl-CoA elongation was also studied over the concentration range 4.5–45 μM [1-¹⁴C]C₁₈CoA. Comparison of the radioactivity incorporated into the fatty acids formed using [2-¹⁴C]malonyl-CoA in the presence of C₁₈CoA, on the one hand, and [1-¹⁴C]C₁₈CoA in the presence of malonyl-CoA, on the other, demonstrated clearly that the acyl chain of the acyl-CoA was elongated by malonyl-CoA.     

Lipids of the Green Alga BotryococcusCultured in a Batch Mode

The lipid fraction of the green alga Botryococcuscultured in a batch mode was found to contain polar lipids (more than 50% of the total lipids), di- and triacylglycerols, sterols and their esters, free fatty acids, and hydrocarbons. In aging culture, the content of polar lipids somewhat decreased and that of triacylglycerols increased by more than four times. The content of hydrocarbons in the algal biomass did not exceed 0.9% and depended little on the culture age. Intracellular lipids contained saturated and unsaturated (mono-, di-, and trienoic) fatty acids. The maximum content of C_{16 : 3}and -C_{18 : 3}fatty acids (up to 35% of the total fatty acids) was detected in the phase of active growth. The extracellular and intracellular lipids of the alga differed in the proportion of particular lipids and in the fatty acid pattern.     

Surface wax of Andreaea and Pogonatum species

The mosses Andreaea rupestris, Pogonatum aloides and P. urnigerum contain surface waxes in amounts of 0.05–0.12% dry wt. The waxes consisted of esters (C₃₈-C₅₄), primary alcohols (C₂₀-C₃₂), free fatty acids (C₁₆-C₃₀), and alkanes (C₂₁-C₃₁). Additionally, aldehydes (C₂₂-C₃₀) were major constituents in the wax of P. urnigerum. The classes and their chain length distributions in the surface waxes of these mosses are comparable to those of epicuticular waxes of higher plants.     

In vitro culture of coconut endosperm: callus induction and its fatty acids

Summary Successful induction of callus from coconut endosperm was achieved by using the tissue situated near the micropylar end of a young fruit. For initiation of callus, a high concentration of auxin (20 to 100 ppm) was added to the basal medium containing activated charcoal. Subcultured callus showed a 40-fold increase during culture of three months. Based on the analysis of fatty acid composition, the maturation of endosperm was characterized by an increase in short chain fatty acids (C₈, C₁₀, C₁₂, C₁₄)and a decrease in long chain fatty acids (C₁₆, C_{18: 1}, C_{18: 2}). In developing endosperms, proportion of short chain fatty acids was higher in lipids of the antipodal than those of other regions. In the final stage of maturation, around 82% of total fatty acids was short chain fatty acids, while the proportion of long chain fatty acids decreased up to 16%. The fatty acid composition of callus subcultured for six months was comparable to that of the immature endosperm. Lipids were accumulated in callus as globular bodies.     

Fatty acids of Pinus elliottii tissues

The total fatty constituents of slash pine (Pinus elliottii) tissue cultures, seeds and seedlings were examined by GLC and MS. Qualitatively, the fatty acid composition of these tissues was very similar to that reported for other pine species. The fatty acid contents of the tissue cultures resembled that of the seedling tissues. In addition to the fatty acids common to botanical materials, Δ⁵-C₁₈ and -C₂₀ nonmethy lene-interrupted polyunsaturated acids were present in low relative abundances. The branched-chain C₁₇ acid reported for several other Pinus species was confirmed as the anteiso isomer.     

Biosynthesis of acyclic methyl branched polyunsaturated hydrocarbons inPseudomonas maltophilia

Summary The hydrocarbon composition ofPseudomonas maltophilia was determined by gas chromatography-mass spectrometry. Mono-, di- and tri-unsaturated alkenes were identified with a predominance of polyunsaturated components. The carbon chains of the alkenes contained single methyl branches iniso andanteiso position and double methyl branches in theiso-iso andanteiso-anteiso configurations. The composition of the hydrocarbons from cells grown in synthetic media enriched with amino acids or volatile fatty acids demonstrated that the probable precursors incorporated into individual hydrocarbons were branched and normal fatty acid chains in the range from C₃ to C₁₆. The probable fatty acid precursors which were connected together to form the major triunsaturated hydrocarbon chains were two monounsaturated chains, whereas the major liunsaturated chains resulted from condensation of saturated and monounsaturated chains. The probable precursors for the major monounsaturated hydrocarbons were C₁₄ (C₁₅) and C₁₆ (C₁₅) fatty acids. The accumulation of hydrocarbons was not detected until the cells were in the late exponential phase of growth; the maximal levels were reached at the mid-stationary phase of growth.     

Specificity effects in the biosynthesis of fatty acids in Bacillus acidocaldarius

Addition to Bacillus acidocaldarius of acids which can act as primers for fatty acid synthesis promote the synthesis of corresponding fatty acids competitively. The effective acids are n?C₅ to -?₇ (not C₄ or C₈), iso- and anteiso-C, and ?C, (not C4), and a range of cyclic acids from cyclobutylacetic and cyclopentanecarboxylic to cycloheptylacetic. New non-natural ω-cyclobutyl-, ω-cyclopentyl-, and ω-cycloheptyl-fatty acids are obtainable. The range of acceptable primers and the range of fatty acids produced therefrom indicate, respectively, the substrate specificities of the transacylase which introduces acyl species into fatty acids synthesis and the one which removes them. The specificity of the primer transacylase may be similar to that in some rumen anaerobes.     

Lipid synthesis by <Emphasis Type="Italic">Geomyces pannorum</Emphasis> under the impact of stress factors

Lipogenic activity and fatty acid composition of two strains of Geomyces pannorum were studied in the course of fungal growth. The strains were isolated from an Arctic cryopeg lens (VKM FW-2241) and from Central Russia (VKM F-3808). The adaptive reactions in both strains towards the temperature decreasing to 2°C involved intensification of the fatty acid desaturation. The degree of lipid unsaturation increased mainly due to a higher amount of α-linolenic acid (α-C_18:3) especially in the case of strain VKM FW-2241. Elevated NaCl concentration in the medium enhanced the level of linoleic acid (C_18:2) which apparently played a specific role in osmoprotection. Strain VKM FW-2241 was more tolerant to increased salinity than strain VKM F-3808. Almost complete inhibition of the growth of strains VKM F-3808 and VKM FW-2241 occurred at salinity of 10 and 20%, respectively; however, the viability of the strains was not affected. Under the combined effect of high salinity and hypothermia, the ratio between C_18:2 and α-C_18:3 acids was intermediate, indicating that these acids were involved in two adaptation mechanisms. The inhibition of fungal growth under stress was found to result in lipid overproduction. An increased pool of energy-rich lipids in fungi possibly contributes to their strategy of cell survival.     

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

The fatty acid composition of cerebrosides, sulphatides and ceramides was determined at 15-16 days post partum in the brain of the Jimpy mutant and in littermate controls. There was a marked deficit in the long chain fatty acids (C₂₂-C₂₄) of cerebrosides and sulphatides of Jimpy brain, with the unsubstituted fatty acids affected more than the alpha-hydroxy fatty acids. A decrease of long chain normal fatty acids was also found in the ceramides of Jimpy brain. The deficit of long chain fatty acids in these sphingolipids of the Jimpy brain was more severe than that found in the Quaking mutant which has a less extensive disorder of myelin formation.     

Possible role of volatile Fatty acids and abscisic Acid in the dormancy of oats

Species of Avena differ markedly in their levels of pre- and post-harvest dormancy. These species offer the opportunity of determining if dormancy is related to the endogenous level of growth inhibitor. Germinability in two species of differing levels of dormancy, common oat Avena sativa L., and wild oat Avena fatua L. was assessed as were the contents of abscisic acid and volatile fatty acids of chain length C₆-C₁₀. In A. sativa which did not possess postharvest dormancy there was no correlation between germination and inhibitor levels but in A. fatua the relationship between the content of fatty acid and dormancy was good. The loss of these fatty acids in dry storage by evaporation could explain after ripening.     

Free fatty acid determination by peroxidase catalysed luminol chemiluminescence

A sensitive, specific, and partly automatic method for the analysis of free fatty acids is described. The assay involves activation of free fatty acids by acyl-CoA synthetase (EC 6.2.1.3) followed by oxidation of the thioesters by acyl-CoA oxidase. The H₂O₂ formed is determined in a reaction catalysed by horseradish peroxidase (EC 1.11.1.7) using luminol as electron donor. The assay has a linear range of 0.05 to 5 nmol of different free fatty acids (C₁₀-C₁₈) in the original sample. The efficiency of the method toward capric, lauric, myristic, palmitic, palmitoleic, stearic, oleic, and linoleic acid measured as recovery of light emission compared to that of H₂O₂ standards, was over 90%. AffiGel 501 was used to covalently bind the free thiol group in CoASH eliminating interference of this substance in the peroxidase-luminol reaction.     

Protein deficiency and age related alterations in rat peritoneal macrophage lipids

The effects of dietary protein restriction and age on the thioglycollate elicited peritoneal macrophage lipid constituents were studied. Impact of subtle changes in lipid components on macrophage functions have been assessed. Lipid profiles of macrophages recovered from rats fed 20 and 4% protein diets and stock diet fed rats (0 and 3 wk) were comparable qualitatively. Quantitative analysis however revealed significant decrease in phospholipids (30–40%) and consequent elevation of cholesterol/phospholipid molar ratios in the protein depleted and young rats (0 wk), compared to the protein fed groups. The protein deficient and the young rats also exhibited accumulation of certain neutral lipids and reduction in triglycerides. Analysis of fatty acid methyl esters of macrophage phospholipids revealed the predominance of long chain polyunsaturated fatty acids even when oleic (C_18:1) and linoleic (C_18:2) formed the bulk of unsaturated fatty acids in the diet. However, the long chain poly unsaturated fatty acid content, particularly the docosahexaenoic acid (C_22:6n-3) was greatly reduced in the protein depleted and 0 wk rats. Observed changes in the long chain polyunsaturated fatty acids of macrophage phospholipids may be of physiological significance as they modulate the immunological functions of the cell.     

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.     

Inhibitory effect of medium-chain-length fatty acids on synthesis of polyhydroxyalkanoates from volatile fatty acids by Ralstonia eutropha

Medium-chain-length fatty acids, such as nonanoic (9:0) and octanoic (8:0) acids, are more toxic to Ralstonia eutropha than volatile fatty acids such as acetic, propionic and butyric acids. Nonanoic acid was degraded to acetic and propionic acids via -oxidation by Ralstonia eutropha for cell growth and synthesis of polyhydroxyalkanoates (PHAs). In a mixture of the fatty acids, utilization of nonanoic acid was depressed by acetic and propionic acids, and vice versa. The PHA accumulation from the volatile fatty acids was decreased from 53% (w/w) of dry cell mass to 23% due to the nonanoic acid. Similar phenomena were also observed with octanoic acid and its metabolic intermediates, acetic and butyric acids.     

Production and characterization of a C15-surfactin-O-methyl ester by a lipopeptide producing strain Bacillus subtilis HSO121

A surfactin mono-methyl ester was isolated from the cell broth of Bacillus subtilis HSO121 by acid precipitation, methanol extraction and two-step chromatographic methods. The structure was analyzed by GC/MS, HPLC, ESI Q-TOF MS/MS and NMR. It indicates that this ester is a C₁₅-surfactin-O-methyl ester of which the fatty acid portions are composed of iso-C₁₅ and anteiso-C₁₅ β-hydroxy fatty acids. Based on the critical micelle concentration (CMC) in phosphate buffered saline (PBS, pH 8.0) and the IC₅₀ value on Hela cell lines, the C₁₅-surfactin-O-methyl ester has a stronger surface activity and lower antitumoral effect than C₁₅-surfactin does. It is found that the Glu residues of surfactin-like lipopeptides play a role in their anti-proliferative effects on Hela cells. The possible antitumoral activity of surfactin is discussed in relation to their structures.     

Further evidence for an elongation-decarboxylation mechanism in the biosynthesis of paraffins in leaves

In isolated tobacco leaves l-valine-U-¹⁴C gave rise to labeled even-numbered isobranched fatty acids containing 16 to 26 carbon atoms and iso C₂₉, iso C₃₁, and iso C₃₃ paraffins. l-Isoleucine-U-¹⁴C on the other hand produced labeled odd-numbered anteiso C₁₇ to C₂₇ fatty acids and anteiso C₃₀ and C₃₂ paraffins. Trichloroacetic acid inhibited the incorporation of isobutyrate into C₂₀ and higher fatty acids and paraffins without affecting the synthesis of the C₁₆ and C₁₈ fatty acids. Thus the very long branched fatty acids are biosynthetically related to the paraffins. In Senecio odoris leaves acetate-1-¹⁴C was incorporated into the paraffins (mainly n-C₃₁) only in the epidermis although acetate was readily incorporated into fatty acids in the mesophyll tissue. Similarly only the epidermal tissue incorporated acetate into fatty acids longer than C₁₈ suggesting that the epidermis is the site of synthesis of both paraffins and the very long fatty acids. In broccoli leaves n-C₁₂ acid labeled with ¹⁴C in the carboxyl carbon and ³H in the methylene carbons was incorporated into C₂₉ paraffin without the loss of ¹⁴C relative to ³H. Since n-C₁₈ acid is known to be incorporated into the paraffin without loss of carboxyl carbon these results suggest that the condensation of C₁₂ acid with C₁₈ acid is not responsible for n-C₂₉ paraffin synthesis in this tissue. Thus all the experimental evidence thus far obtained strongly suggests that elongation of fatty acids followed by decarboxylation is the most likely pathway for paraffin biosynthesis in leaves.