1. Molecular cloning and characterization of an almond 9-hydroperoxide lyase, a new CYP74 targeted to lipid bodies.
- Author
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Mita G, Quarta A, Fasano P, De Paolis A, Di Sansebastiano GP, Perrotta C, Iannacone R, Belfield E, Hughes R, Tsesmetzis N, Casey R, and Santino A
- Subjects
- Amino Acid Sequence, Base Sequence, Cloning, Molecular, DNA Primers, Genomic Library, Hydro-Lyases metabolism, Lipid Metabolism, Molecular Sequence Data, Phylogeny, Recombinant Fusion Proteins metabolism, Recombinant Proteins metabolism, Reverse Transcriptase Polymerase Chain Reaction, Seeds enzymology, Sequence Alignment, Sequence Homology, Amino Acid, Hydro-Lyases genetics, Prunus enzymology
- Abstract
Oxylipin metabolism represents one of many defence mechanisms employed by plants. It begins with the oxygenation of polyunsaturated fatty acids by lipoxygenases to form fatty acid hydroperoxides that are substrates for several enzymes, including specialized cytochrome P450s known as CYP74s. The targeting of a new CYP74, a 9-hydroperoxide lyase (HPL) from almonds, to the endomembrane system and lipid bodies, both as enzyme activity in almond seeds and as GFP fusions transiently expressed in tobacco protoplasts, is described. Such association of a CYP74 with lipid bodies has not been reported previously. Also described are the properties of a 9-HPL gene, the developmental regulation of its expression, the production and characterization of recombinant 9-HPL in Escherichia coli, and the developmental correlation between gene expression, enzyme activity, and the appearance of volatile C9 aldehydes from HPL action.
- Published
- 2005
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