Genetic mechanism of oleocanthal biosynthesis in olive

Abstract

Oleocanthal is an important secondary metabolite that has beenreported to be useful against important human diseases includingcancer. The aim of this study was to identify and characterize thekey biochemical and genetic components of oleocanthal biosyn-thesis. To determine the biochemical components of the pathway,multiple olive cultivars along with their spatial and temporalpoints were determined. The expression levels of multiple candi-date genes were also aimed via real-time PCR. NucleotideBLAST and protein BLAST (for comparison of the similarity ofthe candidate genes with that of other organisms) were conductedon NCBI web page. Phylogenetic tree construction, amino acidcomposition analysis, nucleotide composition analysis, hydropa-thy analysis and translations through ExPASY were conducted.Primer3 was used to design forward and reverse primers toamplify the target genes from different olive tissues at differenttimes. Analysis of the first candiate gene with BioEdit programrevealed that A+T ratio was more than G+C according to thenucleotide composition analysis. According to amino acid com-position analysis isoleucine, lysine and leucine were more thanother amino acids while Kyte&Doolittle hyddropathy analysisrevealed that the protein was hydrophilic. Abundance ofhydrophobic amino acids (leucine and isolecine) along with anabundant hydrophilic amino acid (lysine) suggest the existance ofhydrophobic pockets in the protein which may mean a membranebound protein or a sitoplasmic protein with a strong hydropho-bic core. The molecular weight of the protein was 56 kDa with apI of 9.21. The protein was found to have a signal peptide.According to the SOSUIGramN, intracellular localization wasfound to be in the inner membrane. Analysis of other candidategenes contiunes.Acknowledgements: This study was supported by TUBITAKwith grant number 110O108.