Towards Elucidating Carnosic Acid Biosynthesis in Lamiaceae : Functional Characterization of the Three First Steps of the Pathway in Salvia fruticosa and Rosmarinus officinalis
Dragana Božić,Dimitra Papaefthimiou,Kathleen Brückner,Ric C. H. de Vos,Constantinos A. Tsoleridis,Dimitra Katsarou,Antigoni Papanikolaou,Irini Pateraki,Fani Chatzopoulou,Eleni Dimitriadou,Stefanos Kostas,David Manzano,Ulschan Scheler,Albert Ferrer,Alain Tissier,Antonios M. Makris,Sotirios C. Kampranis,Angelos K. Kanellis +17 more
TLDR
Carnosic acid 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 and Rosmarinus officinalis.Abstract:
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. Recombinant 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 ferruginol synthases, thus revealing the enzymatic activities responsible for the first three steps leading to CA in two Lamiaceae genera.read more
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Terpene Synthases as Metabolic Gatekeepers in the Evolution of Plant Terpenoid Chemical Diversity.
TL;DR: Current knowledge on the functional diversity and molecular evolution of the plant TPS family that underlies the chemical diversity of bioactive terpenoids across the plant kingdom is reviewed.
Journal ArticleDOI
Cytochrome P450 enzymes: A driving force of plant diterpene diversity.
Ulschan Bathe,Alain Tissier +1 more
TL;DR: A phylogenetic analysis showed that only few CYP clans are represented in diterpene biosynthesis, namely CYP71, CYP85 and CYP72, indicating specific expansion of these clades in plant diterpenes biosynthesis.
Journal ArticleDOI
Glandular trichomes: micro-organs with model status?
TL;DR: What novel concepts investigations on glandular trichomes have brought to the field of specialized metabolism, in particular with respect to chemical and enzymatic diversity are reviewed.
Journal ArticleDOI
Carnosic acid and carnosol, two major antioxidants of rosemary, act through different mechanisms
Margot Loussouarn,Anja Krieger-Liszkay,Ljubica Svilar,Antoine Bily,Simona Birtic,Michel Havaux +5 more
TL;DR: In vitro analyses, using high-performance liquid chromatography-ultraviolet and luminescence imaging, revealed that carnosic acid and its major oxidized derivative, carnosol, protect lipids from oxidation, confirming that chemical quenching of ROS by carnosIC acid takes place in planta.
Journal ArticleDOI
Total biosynthesis of the cyclic AMP booster forskolin from Coleus forskohlii
Irini Pateraki,Johan Andersen-Ranberg,Niels Bjerg Jensen,Sileshi Gizachhew Wubshet,Allison M. Heskes,Victor Forman,Björn M. Hallström,Britta Hamberger,Mohammed Saddik Motawia,Carl Erik Olsen,Dan Staerk,Jørgen Hansen,Birger Lindberg Møller,Björn Hamberger +13 more
TL;DR: A minimal set of three P450s in combination with a single acetyl transferase was identified that catalyzes the conversion of 13R-manoyl oxide into forskolin as demonstrated by transient expression in Nicotiana benthamiana.
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