Detection of unprecedented allene oxide synthase member of CYP74B subfamily: CYP74B33 of carrot (Daucus carota).

Enzymes of CYP74 family widespread in higher plants control the metabolism of fatty acid hydroperoxides to numerous bioactive oxylipins. Hydroperoxide lyases (HPLs, synonym: hemiacetal synthases) of CYP74B subfamily belong to the most common CYP74 enzymes. HPLs isomerize the hydroperoxides to the short-lived hemiacetals, which are spontaneously decomposed to aldehydes and aldoacids.

Allene Oxide Synthase Pathway in Cereal Roots: Detection of Novel Oxylipin Graminoxins.

Young roots of wheat, barley, and sorghum, as well as methyl jasmonate pretreated rice seedlings, undergo an unprecedented allene oxide synthase pathway targeted to previously unknown oxylipins -. These Favorskii-type products, (4)-2-pentyl-4-tridecene-1,13-dioic acid (), (2')-2-(2'-octenyl)-decane-1,10-dioic acid (), and (2',5')-2-(2',5'-octadienyl)-decane-1,10-dioic acid (), have a carboxy function at the side chain, as revealed by their MS and NMR spectral data. Compounds - were the major oxylipins detected, along with the related α-ketols.

Identification of CYP443D1 (CYP74 clan) of Nematostella vectensis as a first cnidarian epoxyalcohol synthase and insights into its catalytic mechanism.

The CYP74 clan enzymes are responsible for the biosynthesis of numerous bioactive oxylipins in higher plants, some Proteobacteria, brown and green algae, and Metazoa. A novel putative CYP74 clan gene CYP443D1 of the starlet sea anemone (Nematostella vectensis, Cnidaria) has been cloned, and the properties of the corresponding recombinant protein have been studied in the present work. The recombinant CYP443D1 was incubated with the 9- and 13-hydroperoxides of linoleic and α-linolenic acids (9-HPOD, 13-HPOD, 9-HPOT, and 13-HPOT, respectively), as well as with the 9-hydroperoxide of γ-linolenic acid (γ-9-HPOT) and 15-hydroperoxide of eicosapentaenoic acid (15-HPEPE).

Oxylipins in the spikemoss Selaginella martensii: Detection of divinyl ethers, 12-oxophytodienoic acid and related cyclopentenones.

Green tissues of spikemoss Selaginella martensii Spring possessed the complex oxylipins patterns. Major oxylipins were the products of linoleic and α-linolenic acids metabolism via the sequential action of 13-lipoxygenase and divinyl ether synthase (DES) or allene oxide synthase (AOS). AOS products were represented by 12-oxophytodienoic acid (12-oxo-PDA) isomers.

Stereospecific biosynthesis of (9S,13S)-10-oxo-phytoenoic acid in young maize roots.

Profiling of oxylipins from young maize roots revealed complex patterns of products mainly originating from the combined actions of 9- and 13-lipoxygenases and allene oxide synthase (AOS). A distinctive feature was the high content of the cyclopentenone 10-oxo-11-phytoenoic acid (10-oxo-PEA). Incubations with [1-14C]linoleic acid led to the formation of the α-ketols 13-hydroxy-12-oxo-9-octadecenoic acid and 9-hydroxy-10-oxo-12-octadecenoic acid as well as the cyclopentenones 12-oxo-10-phytoenoic acid (12-oxo-PEA) and 10-oxo-PEA in a ratio of 10:2:1:3.

Sterol binding by methyl-β-cyclodextrin and nystatin--comparative analysis of biochemical and physiological consequences for plants.

The dependence of membrane function on its sterol component has been intensively studied with model lipids and isolated animal membranes, but to a much lesser extent with plant membranes. Depleting membrane sterols could be predicted to have a strong effect on membrane activity and have harmful physiological consequences. In this study, we characterized membrane lipid composition, membrane permeability for ions, some physiological parameters, such as H2O2 accumulation, formation of autophagosomal vacuoles, and expression of peroxidase and autophagic genes, and cell viability in the roots of wheat (Triticum aestivum L.

Isolation and structure elucidation of linolipins C and D, complex oxylipins from flax leaves.

Two complex oxylipins (linolipins C and D) were isolated from the leaves of flax plants inoculated with phytopathogenic bacteria Pectobacterium atrosepticum. Their structures were elucidated based on UV, MS and NMR spectroscopic data. Both oxylipins were identified as digalactosyldiacylglycerol (DGDG) molecular species.

Thermal conversions of fatty acid peroxides to cyclopentenones: a biomimetic model for allene oxide synthase pathway.

The trimethylsilyl (TMS) peroxides of linoleic acid 9(S)-hydroperoxide (TMS or Me esters) were subjected to gas chromatography-mass spectrometry (GC-MS) analyses. The cyclopentenones, trans- and cis-10-oxo-11-phytoenoic acid (10-oxo-PEA, Me or TMS esters) were first time detected as the products of TMS-peroxide thermal conversions. The major products were ketodienes, epoxyalcohols, hemiacetals and decadienals.

Structural details of pectic galactan from the secondary cell walls of flax (Linum usitatissimum L.) phloem fibres.

Details of the backbone and side chain structure of pectic β-(1→4)-galactan from the secondary cell walls of flax phloem fibres were characterised by NMR and mass spectrometry of the fragments obtained after partial hydrolysis with specific endogalactanase and rhamnogalacturonan hydrolase. The proportions of branched and linear rhamnose in the backbone of the polymer equalled 72% and 28%, respectively. Rhamnose branched with a single galactose residue comprised 47% of the total rhamnose; thus, in the bulk of the polymer backbone, rhamnose had 0-1 galactose residues.

Hydroperoxide lyase cascade in pea seedlings: Non-volatile oxylipins and their age and stress dependent alterations.

The profiles of non-volatile oxylipins of pea (Pisum sativum) seedlings were examined by gas chromatography-mass spectrometry after invitro incubation with α-linolenic acid. The 13-lipoxygenase/hydroperoxide lyase (HPL) products were predominant in the leaves, while the roots possess both 13- and 9-HPL products. Allene oxide synthase (AOS) and divinyl ether synthase (DES) products were not detected in the leaves or in the roots of any age.

Unprecedented pathogen-inducible complex oxylipins from flax--linolipins A and B.

Oxylipins constitute a large family of bioregulators, biosynthesized via unsaturated fatty acid oxidation. This study reports the detection of an unprecedented family of complex oxylipins from flax leaves. Two major members of this family, compounds 1 and 2, were isolated and purified.