| Title: | Homologous and heterologous expression of grapevine E-(beta)-caryophyllene synthase (VvGwECar2) |
| Author(s): | Salvagnin U; Carlin S; Angeli S; Vrhovsek U; Anfora G; Malnoy M; Martens S; |
| Address: | "Faculty of Science and Technology, Free University of Bozen-Bolzano, Piazza Universita 5, 39100 Bolzano, Italy; Research and Innovation Centre, Fondazione Edmund Mach, Via E. Mach 1, 38010 S. Michele all'Adige, TN, Italy. Electronic address: umberto.salvagnin@fmach.it. Research and Innovation Centre, Fondazione Edmund Mach, Via E. Mach 1, 38010 S. Michele all'Adige, TN, Italy. Electronic address: silvia.carlin@fmach.it. Faculty of Science and Technology, Free University of Bozen-Bolzano, Piazza Universita 5, 39100 Bolzano, Italy. Electronic address: Sergio.Angeli@unibz.it. Research and Innovation Centre, Fondazione Edmund Mach, Via E. Mach 1, 38010 S. Michele all'Adige, TN, Italy. Electronic address: urska.vrhovsek@fmach.it. Research and Innovation Centre, Fondazione Edmund Mach, Via E. Mach 1, 38010 S. Michele all'Adige, TN, Italy. Electronic address: gianfranco.anfora@fmach.it. Research and Innovation Centre, Fondazione Edmund Mach, Via E. Mach 1, 38010 S. Michele all'Adige, TN, Italy. Electronic address: mickael.malnoy@fmach.it. Research and Innovation Centre, Fondazione Edmund Mach, Via E. Mach 1, 38010 S. Michele all'Adige, TN, Italy. Electronic address: stefan.martens@fmach.it" |
| DOI: | 10.1016/j.phytochem.2016.08.002 |
| ISSN/ISBN: | 1873-3700 (Electronic) 0031-9422 (Linking) |
| Abstract: | "E-(beta)-caryophyllene is a sesquiterpene volatile emitted by plants and involved in many ecological interactions within and among trophic levels and it has a kairomonal activity for many insect species. In grapevine it is a key compound for host-plant recognition by the European grapevine moth, Lobesia botrana, together with other two sesquiterpenes. In grapevine E-(beta)-caryophyllene synthase is coded by the VvGwECar2 gene, although complete characterization of the corresponding protein has not yet been achieved. Here we performed the characterization of the enzyme after heterologous expression in E. coli, which resulted to produce in vitro also minor amounts of the isomer alpha-humulene and of germacrene D. The pH optimum was estimated to be 7.8, and the K(m) and K(cat) values for farnesyl pyrophosphate were 31.4 muM and 0.19 s(-1) respectively. Then, we overexpressed the gene in the cytoplasm of two plant species, Arabidopsis thaliana and the native host Vitis vinifera. In Arabidopsis the enzyme changed the plant head space release, showing a higher selectivity for E-(beta)-caryophyllene, but also the production of thujopsene instead of germacrene D. Overall plants increased the E-(beta)-caryophyllene emission in the headspace collection by 8-fold compared to Col-0 control plants. In grapevine VvGwECar2 overexpression resulted in higher E-(beta)-caryophyllene emissions, although there was no clear correlation between gene activity and sesquiterpene quantity, suggesting a key role by the plant regulation machinery" |
| Keywords: | Alkyl and Aryl Transferases/genetics/*metabolism Arabidopsis/*metabolism Escherichia coli/genetics Monocyclic Sesquiterpenes Polycyclic Sesquiterpenes Polyisoprenyl Phosphates Sesquiterpenes/*chemistry/metabolism Stereoisomerism Vitis/chemistry/enzymology; |
| Notes: | "MedlineSalvagnin, Umberto Carlin, Silvia Angeli, Sergio Vrhovsek, Urska Anfora, Gianfranco Malnoy, Mickael Martens, Stefan eng England 2016/08/27 Phytochemistry. 2016 Nov; 131:76-83. doi: 10.1016/j.phytochem.2016.08.002. Epub 2016 Aug 22" |
