Towards elucidating carnosic acid biosynthesis in Lamiaceae : functional characterization of the three first steps of the pathway in Salvia fruticosa and Rosmarinus officinalis
Božić, Dragana (Aristoteleio Panepistēmio Thessalonikēs. Department of Pharmaceutical Sciences)
Papaefthimiou, Dimitra (Aristoteleio Panepistēmio Thessalonikēs. Department of Pharmaceutical Sciences)
Brückner, Kathleen (Leibniz-Institut für Pflanzenbiochemie)
de Vos, Ric C. H. (Wageningen UR. Plant Breeding)
Tsoleridis, Constantinos A. (Aristoteleio Panepistēmio Thessalonikēs. Department of Chemistry)
Katsarou, Dimitra (Aristoteleio Panepistēmio Thessalonikēs. Department of Pharmaceutical Sciences)
Papanikilaou, Antigoni (Aristoteleio Panepistēmio Thessalonikēs. Department of Pharmaceutical Sciences)
Pateraki, Irini (Centre de Recerca en Agrigenòmica)
Chatzopoulou, Fani M. (Aristoteleio Panepistēmio Thessalonikēs. Department of Pharmaceutical Sciences)
Dimitriadou, Eleni (Aristoteleio Panepistēmio Thessalonikēs. Department of Pharmaceutical Sciences)
Kostas, Stefanos (Aristoteleio Panepistēmio Thessalonikēs. School of Agriculture)
Manzano, David (Centre de Recerca en Agrigenòmica)
Scheler, Ulschan (Leibniz-Institut für Pflanzenbiochemie)
Ferrer, Albert (Centre de Recerca en Agrigenòmica)
Tissier, Alain (Leibniz-Institut für Pflanzenbiochemie)
Makris, Antonios M. (Centre for Research and Technology Hellas (Tessalònica, Grècia))
Kampranis, Sotirios C. (Panepistēmio tēs Krētēs. Department of Biochemistry)
Kanellils, Angelos K. (Aristoteleio Panepistēmio Thessalonikēs. Department of Pharmaceutical Sciences)

Data:	2015
Resum:	Carnosic acid (CA) is a phenolic diterpene with anti-tumour, anti-diabetic, antibacterial and neuroprotective properties that is produced by a number of species from several genera of the Lamiaceae family, including Salvia fruticosa (Cretan sage) and Rosmarinus officinalis (Rosemary). To elucidate CA biosynthesis, glandular trichome transcriptome data of S. fruticosa were mined for terpene synthase genes. Two putative diterpene synthase genes, namely SfCPS and SfKSL, showing similarities to copalyl diphosphate synthase and kaurene synthase-like genes, respectively, were isolated and functionally characterized. Re-combinant expression in Escherichia coli followed by in vitro enzyme activity assays confirmed that SfCPS is a copalyl diphosphate synthase. Coupling of SfCPS with SfKSL, both in vitro and in yeast, resulted in the synthesis miltiradiene, as confirmed by 1D and 2D NMR analyses (1H, 13C, DEPT, COSY H-H, HMQC and HMBC). Coupled transient in vivo assays of SfCPS and SfKSL in Nicotiana benthamiana further confirmed production of miltiradiene in planta. To elucidate the subsequent biosynthetic step, RNA-Seq data of S. fruticosa and R. officinalis were searched for cytochrome P450 (CYP) encoding genes potentially involved in the synthesis of the first phenolic compound in the CA pathway, ferruginol. Three candidate genes were selected, SfFS, RoFS1 and RoFS2. Using yeast and N. benthamiana expression systems, all three where confirmed to be coding for ferruginolsynthases, thus revealing the enzymatic activities responsible for the first three steps leading to CA in two Lamiaceae genera.
Ajuts:	European Commission 227448 European Commission COSTACTION/FA1006
Drets:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Llengua:	Anglès
Document:	Article ; recerca ; Versió publicada
Publicat a:	PloS one, Vol. 10, issue 5 (May 2015) , e0124106, ISSN 1932-6203

DOI: 10.1371/journal.pone.0124106
PMID: 26020634

28 p, 2.5 MB

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