linoleic-acid and crepenynic-acid

Triacylglycerols (TAGs) containing less common fatty acids (FAs) were isolated from the seeds of three plants (Santalum album, Crepis foetida, and Leucas aspera). These FAs had allenic (laballenic acid, Lb) and acetylenic (crepenynic, C; ximenynic acids, Xi) bonds. TAGs were analyzed on reversed-phase and chiral columns. High-resolution tandem mass spectrometry identified TAGs by positive electrospray ionization (ESI

Topics: Alkynes; Chromatography, High Pressure Liquid; Chromatography, Liquid; Chromatography, Reverse-Phase; Fatty Acids; Linoleic Acid; Oleic Acids; Phosphatidylcholines; Seeds; Spectrometry, Mass, Electrospray Ionization; Stereoisomerism; Tandem Mass Spectrometry; Triglycerides

Polyacetylenes (PAs) are bioactive, specialized plant defense compounds produced by some species in the eudicot clade campanulids. Early steps of PA biosynthesis are catalyzed by Fatty Acid Desaturase 2 (

Topics: Alkynes; Campanulaceae; Evolution, Molecular; Fatty Acid Desaturases; Gene Duplication; Linoleic Acid; Multigene Family; Oleic Acids; Phylogeny; Polyacetylene Polymer; Selection, Genetic

Polyacetylenic lipids accumulate in various Apiaceae species after pathogen attack, suggesting that these compounds are naturally occurring pesticides and potentially valuable resources for crop improvement. These compounds also promote human health and slow tumor growth. Even though polyacetylenic lipids were discovered decades ago, the biosynthetic pathway underlying their production is largely unknown. To begin filling this gap and ultimately enable polyacetylene engineering, we studied polyacetylenes and their biosynthesis in the major Apiaceae crop carrot (

Topics: Alkynes; Arabidopsis; Chromatography, Thin Layer; Daucus carota; Diynes; Enzymes; Fatty Acid Desaturases; Fatty Alcohols; Gas Chromatography-Mass Spectrometry; Linoleic Acid; Oleic Acids; Plant Proteins; Plants, Genetically Modified; Polyacetylene Polymer; Saccharomyces cerevisiae

Crepis alpina acetylenase is a variant FAD2 desaturase that catalyses the insertion of a triple bond at the Delta12 position of linoleic acid, forming crepenynic acid in developing seeds. Seeds contain a high level of crepenynic acid but other tissues contain none. Using reverse transcriptase-coupled PCR (RT-PCR), acetylenase transcripts were identified in non-seed C. alpina tissues, which were highest in flower heads. To understand why functional expression of the acetylenase is limited to seeds, genes that affect acetylenase activity by providing substrate (FAD2) or electrons (cytochrome b5), or that compete for substrate (FAD3), were cloned. RT-PCR analysis indicated that the availability of a preferred cytochrome b5 isoform is not a limiting factor. Developing seeds co-express acetylenase and FAD2 isoform 2 (FAD2-2) at high levels. Flower heads co-express FAD2-3 and FAD3 at high levels, and FAD2-2 and acetylenase at moderate levels. FAD2-3 was not expressed in developing seed. Real-time RT-PCR absolute transcript quantitation showed 10(4)-fold higher acetylenase expression in developing seeds than in flower heads. Collectively, the results show that both the acetylenase expression level and the co-expression of other desaturases may contribute to the tissue specificity of crepenynate production. Helianthus annuus contains a Delta12 acetylenase in a polyacetylene biosynthetic pathway, so does not accumulate crepenynate. Real-time RT-PCR analysis showed relatively strong acetylenase expression in young sunflowers. Acetylenase transcription is observed in both species without accumulation of the enzymatic product, crepenynate. Functional expression of acetylenase appears to be affected by competition and collaboration with other enzymes.

Topics: Alkynes; Cloning, Molecular; Crepis; Cytochromes b5; DNA, Plant; Fatty Acid Desaturases; Fatty Acids, Nonesterified; Gene Amplification; Genome, Plant; Helianthus; Linoleic Acid; Molecular Sequence Data; Oleic Acids; Plant Proteins; RNA, Plant; Seeds; Transcription, Genetic

A new octadecatrienoic acid (6.9%), found as a component of Chrysanthemum zawadskii Herb. (Asteraceae) seed oil, was shown to be the hitherto unknown cis,cis,cis-octadeca-3,9,12-trienoic acid. The oil also contained 8.6% of crepenynic acid in addition to the other common FA. The structures of the new unusual FA and other FA were confirmed by chromatographic (TLC, GC), spectroscopic (IR, UV, and NMR), and MS methods by using different chemical derivatizations (preparation of methyl ester, pyrrolidide, picolinyl esters, and dimethyloxazoline derivatives).

Topics: Alkynes; Chromatography, Gas; Chrysanthemum; Fatty Acids; Fatty Acids, Unsaturated; Gas Chromatography-Mass Spectrometry; Linoleic Acid; Molecular Structure; Oleic Acids; Plant Oils; Seeds; Spectroscopy, Fourier Transform Infrared

Acetylenic bonds are present in more than 600 naturally occurring compounds. Plant enzymes that catalyze the formation of the Delta12 acetylenic bond in 9-octadecen-12-ynoic acid and the Delta12 epoxy group in 12,13-epoxy-9-octadecenoic acid were characterized, and two genes, similar in sequence, were cloned. When these complementary DNAs were expressed in Arabidopsis thaliana, the content of acetylenic or epoxidated fatty acids in the seeds increased from 0 to 25 or 15 percent, respectively. Both enzymes have characteristics similar to the membrane proteins containing non-heme iron that have histidine-rich motifs.

Topics: Acetylene; Alkynes; Amino Acid Sequence; Arabidopsis; Asteraceae; Catalysis; Cloning, Molecular; DNA, Complementary; Epoxy Compounds; Fatty Acid Desaturases; Genes, Plant; Iron; Linoleic Acid; Microsomes; Molecular Sequence Data; NAD; NADP; Oleic Acids; Oxidoreductases; Plant Proteins; Plants, Genetically Modified; Saccharomyces cerevisiae; Seeds; Sequence Alignment

Triple bond analogues of natural fatty acids irreversibly inactivate lipoxygenase during their enzymatic conversion [Nieuwenhuizen, W. F., et al. (1995) Biochemistry 34, 10538-10545]. To gain insight into the mechanism of the irreversible inactivation of soybean lipoxygenase-1, we studied the enzymatic conversion of two linoleic acid analogues, 9(Z)-octadec-9-en-12-ynoic acid (9-ODEYA) and 12(Z)-octadec-12-en-9-ynoic acid (12-ODEYA). During the inactivation process, Fe(III)-lipoxygenase converts 9-ODEYA into three products, i.e. 11-oxooctadec-9-en-12-ynoic acid, racemic 9-hydroxy-10(E)-octadec-10-en-12-ynoic acid, and racemic 9-hydroperoxy-10(E)-octadec-10-en-12-ynoic acid. Fe(II)-lipoxygenase does not convert the inhibitor and is not inactivated by 9-ODEYA. Fe(III)-lipoxygenase converts 12-ODEYA into 13-hydroperoxy-11(Z)-octadec-11-en-9-ynoic acid (34/66 R/S), 13-hydroperoxy11(E)-octadec-11-en-9-ynoic acid (36/64 R/S), 11-hydroperoxyoctadec-12-en-9-ynoic acid (11-HP-12-ODEYA, enantiomeric composition of 33/67), and 11-oxooctadec-12-en-9-ynoic acid (11-oxo-12-ODEYA) during the inactivation process. Also, Fe(II)-lipoxygenase is inactivated by 12-ODEYA. It converts the inhibitor into the same products as Fe(III)-lipoxygenase does, but two additional products are formed, viz. 13-oxo-11(E)-octadec-11-en-9-ynoic acid and 13-oxo-11(Z)-octadec-11-en-9-ynoic acid. The purified reaction products were tested for their lipoxygenase inhibitory activities. The oxo compounds, formed in the reaction of 9-ODEYA and 12-ODEYA, do not inhibit Fe(II)- or Fe(III)-lipoxygenase. The 9- and 13-hydroperoxide products that are formed from 9-ODEYA and 12-ODEYA, respectively, oxidize Fe(II)-lipoxygenase to its Fe(III) state and are weak lipoxygenase inhibitors. 11-HP-12-ODEYA is, however, the most powerful inhibitor and is able to oxidize Fe(II)-lipoxygenase to Fe(III)-lipoxygenase. 11-HP-12-ODEYA is converted into 11-oxo-12-ODEYA by Fe(III)-lipoxygenase. We propose a mechanism for the latter reaction in which Fe(III)-lipoxygenase abstracts the bisallylic hydrogen H-11 from 11-HP-12-ODEYA, yielding a hydroperoxyl radical which is subsequently cleaved into 11-oxo-ODEYA and a hydroxyl radical which may inactivate the enzyme.

Topics: Alkynes; Chromatography, High Pressure Liquid; Ferric Compounds; Ferrous Compounds; Gas Chromatography-Mass Spectrometry; Glycine max; Hydrogen Peroxide; Isomerism; Linoleic Acid; Linoleic Acids; Lipid Peroxidation; Lipoxygenase; Lipoxygenase Inhibitors; Oleic Acids; Quantum Theory; Spectrophotometry, Ultraviolet

The seed oil of the plant Ixiolaena brevicompta is a rich source of crepenynic acid (octadec-cis-9-en-12-ynoic acid), which has been linked with extensive sheep mortalities in Western New South Wales and Queensland, Australia. A number of acetylenic fatty acids have been found to interfere with lipid and fatty acid metabolism and inhibit cyclooxygenase and lipoxygenase enzymes in a variety of tissues. We have investigated the effects of crepenynic acid and ximenynic acid (octadec-trans-11-en-9-ynoic acid) on leukotriene B4 and thromboxane B2 production in rat peritoneal leukocytes and compare them with non-acetylenic compounds linoleic and ricinoleic acids. In concentrations ranging from 10 to 100 microM linoleic acid and ricinoleic acid had only minimal effects on leukotriene B4 and thromboxane B2 production in ionophore-stimulated cells. Ximenynic acid gave dose-dependent inhibition of leukotriene B4, thromboxane B2 and 6-ketoprostaglandin F1 alpha production. Ximenynic acid appears to be a more effective inhibitor of leukotriene B4 than crepenynic acid with an IC50 of 60 microM compared to 85 microM. On the other hand, crepenynic acid is a much more effective inhibitor of the cyclooxygenase products, having an IC50 for thromboxane B2 of less than 10 microM. Both acetylenic fatty acids inhibited phospholipase activity in these cells by 40-50% at a concentration of 100 microM but had no inhibitory effect at 10 microM. These results indicate that crepenynic acid and ximenynic acid differentially inhibit the cyclooxygenase and lipoxygenase products of stimulated leukocytes, and that at high doses of these fatty acids the effect on these products may be partially due to inhibition of phospholipase A2.

Topics: Alkynes; Animals; Cyclooxygenase Inhibitors; Leukocytes; Leukotriene B4; Linoleic Acid; Linoleic Acids; Lipoxygenase Inhibitors; Oleic Acids; Peritoneal Cavity; Phospholipases; Rats; Ricinoleic Acids; Thromboxane B2

Topics: Alkynes; Chromatography, High Pressure Liquid; Depression, Chemical; Esters; Fatty Acids, Essential; Linoleic Acid; Oleic Acids; Plants, Toxic; Solvents