Methyl Jasmonate Induces Lauric Acid ω-Hydroxylase Activity and Accumulation of CYP94A1 Transcripts but Does Not Affect Epoxide Hydrolase Activities in<i>Vicia sativa</i>Seedlings

Pinot, Franck; Benveniste, Irène; Salaün, Jean‐Pierre; Durst, Francis

doi:10.1104/pp.118.4.1481

Cited by 34 publications

(16 citation statements)

References 37 publications

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“…These FA derivatives are produced by CYP94A1 from V. sativa suggesting a potential role in plant defence for this enzyme. This hypothesis receives support from experiments showing that treatment of V. sativa seedlings with the stress hormone methyl jasmonate enhanced CYP94A1 at the transcriptional level [44]. Interestingly, clofibrate treatment also enhanced CYP94A1 at the transcriptional level [13] and increased the proliferation of peroxisomes [45] which have an important role in responses to pathogens [46].…”

Section: Plant Defencementioning

confidence: 97%

Cytochrome P450 metabolizing fatty acids in plants: characterization and physiological roles

2010

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In plants, fatty acids (FA) are subjected to various types of oxygenation reactions. Products include hydroxyacids, as well as hydroperoxides, epoxides, aldehydes, ketones and α,ω‐diacids. Many of these reactions are catalysed by cytochrome P450s (P450s), which represent one of the largest superfamilies of proteins in plants. The existence of P450‐type metabolizing FA enzymes in plants was established approximately four decades ago in studies on the biosynthesis of lipid polyesters. Biochemical investigations have highlighted two major characteristics of P450s acting on FAs: (a) they can be inhibited by FA analogues carrying an acetylenic function, and (b) they can be enhanced by biotic and abiotic stress at the transcriptional level. Based on these properties, P450s capable of producing oxidized FA have been identified and characterized from various plant species. Until recently, the vast majority of characterized P450s acting on FAs belonged to the CYP86 and CYP94 families. In the past five years, rapid progress in the characterization of mutants in the model plant Arabidopsis thaliana has allowed the identification of such enzymes in many other P450 families (i.e. CYP703, CYP704, CYP709, CYP77, CYP74). The presence in a single species of distinct enzymes characterized by their own regulation and catalytic properties raised the question of their physiological meaning. Functional studies in A. thaliana have demonstrated the involvement of FA hydroxylases in the synthesis of the protective biopolymers cutin, suberin and sporopollenin. In addition, several lines of evidence discussed in this minireview are consistent with P450s metabolizing FAs in many aspects of plant biology, such as defence against pathogens and herbivores, development, catabolism or reproduction.

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Section: Plant Defencementioning

confidence: 97%

Cytochrome P450 metabolizing fatty acids in plants: characterization and physiological roles

2010

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“…Furthermore they are also potent elicitors of defense mechanisms (12,13). The hypothesis of CYP94A1 participation in a plant defense event received support from the observation that this enzyme is induced at the transcriptional level by the stress hormone methyl jasmonate (14). Plant fatty acid hydroxylases could also be involved in catabolism like members of the CYP4 family in animals (15,16) or like CYP52A3 and CYP52A4, which enable Candida maltosa to use aliphatic hydrocarbon as a source of carbon and energy (17).…”

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confidence: 99%

Cloning, Functional Expression, and Characterization of CYP709C1, the First Sub-terminal Hydroxylase of Long Chain Fatty Acid in Plants

Kandel

Morant²,

Benveniste

et al. 2005

Journal of Biological Chemistry

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We cloned and characterized CYP709C1, a new plant cytochrome P450 belonging to the P450 family, that so far has no identified function except for clustering with a fatty acid metabolizing clade of P450 enzymes. We showed here that CYP709C1 is capable of hydroxylating fatty acids at the -1 and -2 positions. This work was performed after recoding and heterologous expression of a fulllength cDNA isolated from a wheat cDNA library in an engineered yeast strain. Investigation on substrate specificity indicates that CYP709C1 metabolizes different fatty acids varying in their chain length (C12 to C18) and unsaturation. CYP709C1 is the first identified plant cytochrome P450 that can catalyze sub-terminal hydroxylation of C18 fatty acids. cis-9,10-Epoxystearic acid is metabolized with the highest efficiency, i.e. K m(app) of 8 M and V max(app) of 328 nmol/min/nmol P450. This, together with the fact that wheat possesses a microsomal peroxygenase able to synthesize this compound from oleic acid, strongly suggests that it is a physiological substrate. Hydroxylated fatty acids are implicated in plant defense events. We postulated that CYP709C1 could be involved in plant defense by producing such compounds. This receives support from the observation that (i) sub-terminal hydroxylation of 9,10-epoxystearic acid is induced (15-fold after 3 h) in microsomes of wheat seedlings treated with the stress hormone methyl jasmonate and (ii) CYP709C1 is enhanced at the transcriptional level by this treatment. CYP709C1 transcript also accumulated after treatment with a combination of the safener naphthalic acid anhydride and phenobarbital. This indicates a possible detoxifying function for CYP709C1 that we discussed.

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“…The BNSEH1 clone was isolated from a cDNA library prepared from MeJa‐treated leaves. sEH has been described to be up‐regulated at the transcript level by MeJa in potato [11] but in Vicia sativa seedlings sEH activity remained unaffected by MeJa treatment [32]. No MeJa induction of sEH transcripts was observed in B. napus seedlings kept in hydroponic cultures (S. Bellevik, F. Sitbon & J. Meier) suggesting that BNSEH1 probably has a constitutive expression.…”

Section: Discussionmentioning

confidence: 99%

Brassica napus soluble epoxide hydrolase (BNSEH1)

Bellevik¹,

Zhang²,

Meijer³

2002

European Journal of Biochemistry

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Epoxide hydrolase (EC 3.3.2.3) in plants is involved in the metabolism of epoxy fatty acids and in mediating defence responses. We report the cloning of a full-length epoxide hydrolase cDNA (BNSEH1) from oilseed rape (Brassica napus) obtained by screening of a cDNA library prepared from methyl jasmonate induced leaf tissue, and the 5¢-RACE technique. The cDNA encodes a soluble protein containing 318 amino acid residues. The identity on the protein level is 85% to an Arabidopsis soluble epoxide hydrolase (sEH) and 50-60% to sEHs cloned from other plants. A 5 · His tag was added to the N-terminus of the BNSEH1 and the construct was over-expressed in the yeast Pichia pastoris. The recombinant protein was recovered at high levels after Ni-agarose chromatography of lysed cell extracts, had a molecular mass of 37 kDa on SDS/PAGE and cross-reacted on Western blots with antibodies raised to a sEH from Arabidopsis thaliana. BNSEH1 was shown to be a monomer by gel filtration analysis. The activity was low towards cisstilbene oxide but much higher using trans-stilbene oxide as substrate with V max of 0.47 lmolAEminAEmg . The optimum temperature of the recombinant enzyme was 55°C and the optimum pH 6-7 for trans-stilbene oxide hydrolysis. The isolation of BNSEH1 will facilitate metabolic engineering of epoxy fatty acid metabolism for functional studies of resistance and seed oil modification in this important oilcrop.Keywords: epoxide hydrolase, oilseed rape, yeast, recombinant enzyme, stilbene oxide.Epoxide hydrolases [1] are hydrolytic enzymes that have been found in mammals, plants, yeast, fungi, insects and bacteria. In mammals several forms exist with a heterogeneous tissue distribution of which microsomal and soluble (sEH) epoxide hydrolases have been most extensively studied. These enzymes differ from each other in several aspects such as substrate preference, turnover rate, sensitivity to inhibitors, pH optimum, etc. [2]. Several geometrically different but related epoxides such as cis-stilbene oxide (CSO) and trans-stilbene oxide (TSO) have been found to be useful substrates in order to distinguish soluble from membrane bound epoxide hydrolases [3,4]. These substrate pairs can be applied to crude extracts to assess the relative contribution of membrane bound and soluble forms to the total epoxide hydrolase activity in many species.Based on sequence homology analysis epoxide hydrolase was classified as a member of a super family of hydrolytic enzymes including esterases and lipases, united by an a/b hydrolase fold and a similar catalytic triad motif [5]. The three-dimensional structure of epoxide hydrolase has been resolved for mouse [6], fungal [7] and bacterial enzymes [8]. These studies have confirmed the predicted a/b fold structure, provided a detailed picture of the active site and proposed a mechanism of the catalytic reaction supported by site-directed mutagenesis. Epoxide hydrolases act through a two-step mechanism in which an acidic nucleophile attacks the epoxide ring, forming a covalent intermediate, whi...

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Methyl Jasmonate Induces Lauric Acid ω-Hydroxylase Activity and Accumulation of CYP94A1 Transcripts but Does Not Affect Epoxide Hydrolase Activities inVicia sativaSeedlings

Cited by 34 publications

References 37 publications

Cytochrome P450 metabolizing fatty acids in plants: characterization and physiological roles

Cytochrome P450 metabolizing fatty acids in plants: characterization and physiological roles

Cloning, Functional Expression, and Characterization of CYP709C1, the First Sub-terminal Hydroxylase of Long Chain Fatty Acid in Plants

Brassica napus soluble epoxide hydrolase (BNSEH1)

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