20.500.12556/DiRROS-21607 Reconstitution of the costunolide biosynthetic pathway in yeast and Nicotiana benthamiana The sesquiterpene costunolide has a broad range of biological activities and is the parent compound for many other biologically active sesquiterpenes such as parthenolide. Two enzymes of the pathway leading to costunolide have been previously characterized: germacrene A synthase (GAS) and germacrene A oxidase(GAO), which together catalyse the biosynthesis of germacra-1(10),4,11(13)-trien-12-oic acid. However, the gene responsible for the last step toward costunolide has not been characterized until now. Here weshow that chicory costunolide synthase (CiCOS), CYP71BL3, can catalyse the oxidation of germacra-1(10),4,11(13)-trien-12-oic acid to yield costunolide. Co-expression of feverfew GAS (TpGAS), chicory GAO (CiGAO), and chicory COS (CiCOS) in yeast resulted in the biosynthesis of costunolide. The catalytic activity of TpGAS, CiGAO and CiCOS was also verified in planta by transient expression in Nicotiana benthamiana. Mitochondrial targeting of TpGAS resultedin a significant increase in the production of germacrene A compared with the native cytosolic targeting. When the N. benthamiana leaves were co-infiltrated with TpGAS and CiGAO, germacrene A almost completely disappeared as a result of the presence of CiGAO. Transient expression of TpGAS, CiGAO and CiCOS in N. benthamiana leaves resulted in costunolide production of up to 60 ng.g-1 FW. In addition, two new compounds were formed that were identified as costunolide-glutathione and costunolide-cysteine conjugates. true false true Angleški jezik Ni določen Neznano 2025-03-04 10:02:28 2025-03-04 10:02:28 2025-03-05 03:43:58 0000-00-00 00:00:00 2011 0 0 BSDOCID160498; str. e23255 [12 str.] iss. 8 Vol. 6 2011 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 2011-08-15 577 1932-6203 10.1371/journal.pone.0023255 29051097 RAZ_Liu_Qing_2011.pdf RAZ_Liu_Qing_2011.pdf 1 EFA7B357F6E708D8B220DD7A3B4B84CA 4d3dc2834707a94adf32f446f3c8dd186d9c02c56fd92fe5335c5c0f5777682e 79cdecdc-f8d7-11ef-9f1e-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=30932 https://doi.org/10.1371/journal.pone.0023255 0 83e25c05-f8d7-11ef-9f1e-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=30933 Nacionalni inštitut za biologijo 0 0 0