Cloning and functional characterization of CYP94A2, a medium chain fatty acid hydroxylase from Vicia sativa.

@article{LeBouquin1999CloningAF,
  title={Cloning and functional characterization of CYP94A2, a medium chain fatty acid hydroxylase from Vicia sativa.},
  author={R Le Bouquin and Franck Pinot and Irène Benveniste and J P Sala{\"u}n and Francis Durst},
  journal={Biochemical and biophysical research communications},
  year={1999},
  volume={261 1},
  pages={
          156-62
        }
}
A full length cDNA encoding a new cytochrome P450-dependent fatty acid hydroxylase (CYP94A2) was isolated from a Vicia sativa library. CYP94A2 displays 58% sequence identity with CYP94A1, a fatty acid omega-hydroxylase isolated from the same material. Heterologous expression of CYP94A2 in Saccharomyces cerevisiae yeast strain WAT11 shows that it catalyses the hydroxylation of myristic (C14) acid with a K(m(app)) of 4.0 microM and a turnover rate number of 80 min(-1). In addition, lauric (C12… 
A conservative amino acid substitution alters the regiospecificity of CYP94A2, a fatty acid hydroxylase from the plant Vicia sativa.
TLDR
The observed shift from omega toward omega-1 hydroxylation was prominent with lauric acid as substrate and declined with increasing fatty acid chain length, and a conservative amino acid substitution at that position markedly altered the regiospecificity of CYP94A2.
Molecular Cloning of CYP76B9, a Cytochrome P450 from Petunia hybrida, Catalyzing the ω-Hydroxylation of Capric Acid and Lauric Acid
TLDR
It is found that the ω-hydroxy metabolites of capric acid and lauric acid can affect the plant growth of Arabidopsis thaliana and be affected by preferential expression of the CYP76B9 gene in flowers and leaves.
CYP94A5, a new cytochrome P450 from Nicotiana tabacum is able to catalyze the oxidation of fatty acids to the omega-alcohol and to the corresponding diacid.
TLDR
A cytochrome P450 able to catalyze the complete set of reactions oxidizing a terminal methyl group to the corresponding carboxyl is described, which is enantioselective: after incubation of a synthetic racemic mixture of 9,10-epoxystearic acid, the chirality of the residual epoxide was 40/60 in favor of 9R,10S enantiomer.
Molecular definitions of fatty acid hydroxylases in Arabidopsis thaliana
TLDR
Comparison between these five P450s at constant P450 content over a range of concentrations for individual fatty acids indicates that binding of different fatty acids to CYP86A2 always results in a higher proportion of high spin state heme than binding titrations conducted with CYP84A1 or CYP85A4.
CYP94A1, a plant cytochrome P450-catalyzing fatty acid ω-hydroxylase, is selectively induced by chemical stress in Vicia sativa seedlings
TLDR
The expression of CYP94A1 under these conditions and the concomitant morphological and cytological modifications would suggest the implication of this P450 in a process of plant defense against chemical injury.
Differential Expression and Evolution of the Arabidopsis CYP86A Subfamily1[w]
TLDR
Analysis of the promoter sequences for each of the CYP86A and CYP94B genes with their expression patterns has highlighted a number of elements in current databases that potentially correlate with the responses of individual genes.
Functional analysis of the LACERATA gene of Arabidopsis provides evidence for different roles of fatty acid ω-hydroxylation in development
TLDR
It is indicated that LCR-dependent ω-hydroxylation of fatty acids could be implicated in the biosynthesis of cutin in the epidermis and in preventing postgenital organ fusions.
Overexpression of a novel cytochrome P450 monooxygenase gene, CYP704B1, from Panax ginseng increase biomass of reproductive tissues in transgenic Arabidopsis
TLDR
The results suggest that PgCYP704B1 has a conserved role during male reproduction for fatty acid biosynthesis and its overexpression increases cutin monomers in siliques that eventually could be used for seed production.
Cytochrome P450 Family Member CYP704B2 Catalyzes the ω -Hydroxylation of Fatty Acids and Is Required for Anther Cutin Biosynthesis and Pollen Exine Formation in Rice[W][OA]
TLDR
This study indicates that the ω -hydroxylation pathway of fatty acids relying on this ancient CYP704B family, conserved from moss to angiosperms, is essential for the formation of both cuticle and exine during plant male reproductive and spore development.
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References

SHOWING 1-10 OF 29 REFERENCES
Cloning, Expression in Yeast, and Functional Characterization of CYP81B1, a Plant Cytochrome P450 That Catalyzes In-chain Hydroxylation of Fatty Acids*
TLDR
A systematic exploration of its function revealed that it specifically catalyzes the hydroxylation of medium chain saturated fatty acids, capric, lauric, and myristic acids.
Functional expression in yeast and characterization of a clofibrate-inducible plant cytochrome P-450 (CYP94A1) involved in cutin monomers synthesis.
TLDR
The chemical tagging of a cytochrome P-450-dependent lauric acid omega-hydroxylase from clofibrate-treated Vicia sativa seedlings with [1-14C]11-dodecynoic acid allowed the isolation of a full-length cDNA designated CYP94A1, which was strong regioselectivity shifted with sulphur-containing substrates.
An oleate 12-hydroxylase from Ricinus communis L. is a fatty acyl desaturase homolog.
TLDR
It is suggested that fatty acyl desaturases and hydroxylases share similar reaction mechanisms and provide an example of enzyme evolution.
CYP86A1 from Arabidopsis thaliana encodes a cytochrome P450-dependent fatty acid omega-hydroxylase.
TLDR
Genomic organization analyzed by Southern blot suggested a single gene encoding CYP86A1 in A. thaliana, which was found to catalyze the omega-hydroxylation of saturated and unsaturated fatty acids with chain lengths from C12 to C18 but not of hexadecane.
Methyl jasmonate induces lauric acid omega-hydroxylase activity and accumulation of CYP94A1 transcripts but does not affect epoxide hydrolase activities in vicia sativa seedlings
TLDR
Treatment of etiolated Vicia sativa seedlings by the plant hormone methyl jasmonate (MetJA) led to an increase of cytochrome P450 content and transcripts coding for CYP94A1, a fatty acid omega-hydroxylase, had already accumulated after 1 h of exposure to MetJA.
New cytochrome P450-dependent reactions from wheat: terminal and sub-terminal hydroxylation of oleic acid by microsomes from naphthalic acid anhydride and phenobarbital induced wheat seedlings.
TLDR
The results suggest that at least two different P450 enzymes are involved in the oxidation of oleic and lauric acids in wheat.
The cytochrome P450 4 (CYP4) family.
  • A. Simpson
  • Biology, Chemistry
    General pharmacology
  • 1997
Induction and specificity of a (cytochrome P-450)-dependent laurate in-chain-hydroxylase from higher plant microsomes.
TLDR
The substrate and product specifities of the (cytochrome P-450)-dependent laurate monooxygenase from tuber tissues of Jerusalem artichoke were investigated and Regulation of l Laurate hydroxylase was clearly independent from that of cinnamic acid 4-hydroxyl enzyme.
Involvement of cytochrome P450 2E1 in the (omega-1)-hydroxylation of oleic acid in human and rat liver microsomes.
TLDR
It can be ascertained that P450 2E1 is the main enzyme involved in the (omega-1)-hydroxylation of oleic acid, which was shown to be mainly catalyzed by P450 4A enzymes in human liver microsomes.
...
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