| Title: | Reconstitution of the costunolide biosynthetic pathway in yeast and Nicotiana benthamiana |
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| Authors: | ID Liu, Qing (Author)
ID Majdi, Mohammad (Author)
ID Cankar, Katarina (Author)
ID Goedbloed, Miriam (Author)
ID Charnikhova, Tatsiana (Author)
ID Verstappen, Francel W. A. (Author)
ID de Vos, Ric (Author)
ID Beekwilder, Jules (Author)
ID van der Krol, Sander (Author)
ID Bouwmeester, Harro (Author) |
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| Files: |  PDF - Presentation file, download (553,05 KB)
MD5: EFA7B357F6E708D8B220DD7A3B4B84CA
 URL - Source URL, visit https://doi.org/10.1371/journal.pone.0023255
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| Language: | English |
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| Typology: | 1.01 - Original Scientific Article |
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| Organization: |  NIB - National Institute of Biology
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| Abstract: | 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. |
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| Publication status: | Published |
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| Publication version: | Version of Record |
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| Publication date: | 15.08.2011 |
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| Year of publishing: | 2011 |
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| Number of pages: | str. e23255 [12 str.] |
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| Numbering: | Vol. 6, iss. 8 |
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| PID: | 20.500.12556/DiRROS-21607  |
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| UDC: | 577 |
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| ISSN on article: | 1932-6203 |
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| DOI: | 10.1371/journal.pone.0023255 |
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| COBISS.SI-ID: | 29051097 |
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| Note: | BSDOCID160498;
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| Publication date in DiRROS: | 04.03.2025 |
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| Views: | 616 |
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| Downloads: | 300 |
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