Molecular cloning and characterization of an almond 9-hydroperoxide lyase, a new CYP74 targeted to lipid bodies.
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.
Mita
Giovanni
G
Institute of Sciences of Food Production CNR Section of Lecce, via Monteroni, I-73100 Italy.
Quarta
Angela
A
Fasano
Pasqua
P
De Paolis
Angelo
A
Di Sansebastiano
Gian Pietro
GP
Perrotta
Carla
C
Iannacone
Rina
R
Belfield
Eric
E
Hughes
Richard
R
Tsesmetzis
Nicolas
N
Casey
Rod
R
Santino
Angelo
A
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D015698:Genomic Library; D006836:Hydro-Lyases / Q000235:genetics* / Q000378:metabolism; D050356:Lipid Metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D027861:Prunus / Q000201:enzymology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012639:Seeds / Q000201:enzymology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C443399:9-hydroperoxide lyase; D006836:Hydro-Lyases