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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...

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Autores principales: Liu, Qing, Majdi, Mohammad, Cankar, Katarina, Goedbloed, Miriam, Charnikhova, Tatsiana, Verstappen, Francel W. A., de Vos, Ric C. H., Beekwilder, Jules, van der Krol, Sander, Bouwmeester, Harro J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3156125/
https://www.ncbi.nlm.nih.gov/pubmed/21858047
http://dx.doi.org/10.1371/journal.pone.0023255
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author Liu, Qing
Majdi, Mohammad
Cankar, Katarina
Goedbloed, Miriam
Charnikhova, Tatsiana
Verstappen, Francel W. A.
de Vos, Ric C. H.
Beekwilder, Jules
van der Krol, Sander
Bouwmeester, Harro J.
author_facet Liu, Qing
Majdi, Mohammad
Cankar, Katarina
Goedbloed, Miriam
Charnikhova, Tatsiana
Verstappen, Francel W. A.
de Vos, Ric C. H.
Beekwilder, Jules
van der Krol, Sander
Bouwmeester, Harro J.
author_sort Liu, Qing
collection PubMed
description 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 we show 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 resulted in 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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spelling pubmed-31561252011-08-19 Reconstitution of the Costunolide Biosynthetic Pathway in Yeast and Nicotiana benthamiana Liu, Qing Majdi, Mohammad Cankar, Katarina Goedbloed, Miriam Charnikhova, Tatsiana Verstappen, Francel W. A. de Vos, Ric C. H. Beekwilder, Jules van der Krol, Sander Bouwmeester, Harro J. PLoS One Research Article 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 we show 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 resulted in 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. Public Library of Science 2011-08-15 /pmc/articles/PMC3156125/ /pubmed/21858047 http://dx.doi.org/10.1371/journal.pone.0023255 Text en Liu et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Liu, Qing
Majdi, Mohammad
Cankar, Katarina
Goedbloed, Miriam
Charnikhova, Tatsiana
Verstappen, Francel W. A.
de Vos, Ric C. H.
Beekwilder, Jules
van der Krol, Sander
Bouwmeester, Harro J.
Reconstitution of the Costunolide Biosynthetic Pathway in Yeast and Nicotiana benthamiana
title Reconstitution of the Costunolide Biosynthetic Pathway in Yeast and Nicotiana benthamiana
title_full Reconstitution of the Costunolide Biosynthetic Pathway in Yeast and Nicotiana benthamiana
title_fullStr Reconstitution of the Costunolide Biosynthetic Pathway in Yeast and Nicotiana benthamiana
title_full_unstemmed Reconstitution of the Costunolide Biosynthetic Pathway in Yeast and Nicotiana benthamiana
title_short Reconstitution of the Costunolide Biosynthetic Pathway in Yeast and Nicotiana benthamiana
title_sort reconstitution of the costunolide biosynthetic pathway in yeast and nicotiana benthamiana
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3156125/
https://www.ncbi.nlm.nih.gov/pubmed/21858047
http://dx.doi.org/10.1371/journal.pone.0023255
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