Nicotiana benthamiana as a production platform for artemisinin precursors.

<h4>Background</h4>Production of pharmaceuticals in plants provides an alternative for chemical synthesis, fermentation or natural sources. Nicotiana benthamiana is deployed at commercial scale for production of therapeutic proteins. Here the potential of this plant is explored for rapid...

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Autores principales:	Teun W J M van Herpen, Katarina Cankar, Marilise Nogueira, Dirk Bosch, Harro J Bouwmeester, Jules Beekwilder
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2010
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d0864dd6e0444e0b95ce420dd69c3763
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spelling	oai:doaj.org-article:d0864dd6e0444e0b95ce420dd69c37632021-11-18T07:02:04ZNicotiana benthamiana as a production platform for artemisinin precursors.1932-620310.1371/journal.pone.0014222https://doaj.org/article/d0864dd6e0444e0b95ce420dd69c37632010-12-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21151979/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Production of pharmaceuticals in plants provides an alternative for chemical synthesis, fermentation or natural sources. Nicotiana benthamiana is deployed at commercial scale for production of therapeutic proteins. Here the potential of this plant is explored for rapid production of precursors of artemisinin, a sesquiterpenoid compound that is used for malaria treatment.<h4>Methodology/principal findings</h4>Biosynthetic genes leading to artemisinic acid, a precursor of artemisinin, were combined and expressed in N. benthamiana by agro-infiltration. The first committed precursor of artemisinin, amorpha-4,11-diene, was produced upon infiltration of a construct containing amorpha-4,11-diene synthase, accompanied by 3-hydroxy-3-methylglutaryl-CoA reductase and farnesyl diphosphate synthase. Amorpha-4,11-diene was detected both in extracts and in the headspace of the N. benthamiana leaves. When the amorphadiene oxidase CYP71AV1 was co-infiltrated with the amorphadiene-synthesizing construct, the amorpha-4,11-diene levels strongly decreased, suggesting it was oxidized. Surprisingly, no anticipated oxidation products, such as artemisinic acid, were detected upon GC-MS analysis. However, analysis of leaf extracts with a non-targeted metabolomics approach, using LC-QTOF-MS, revealed the presence of another compound, which was identified as artemisinic acid-12-β-diglucoside. This compound accumulated to 39.5 mg x kg(-1) fwt. Apparently the product of the heterologous pathway that was introduced, artemisinic acid, is further metabolized efficiently by glycosyl transferases that are endogenous to N. benthamiana.<h4>Conclusion/significance</h4>This work shows that agroinfiltration of N. bentamiana can be used as a model to study the production of sesquiterpenoid pharmaceutical compounds. The interaction between the ectopically introduced pathway and the endogenous metabolism of the plant is discussed.Teun W J M van HerpenKatarina CankarMarilise NogueiraDirk BoschHarro J BouwmeesterJules BeekwilderPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 5, Iss 12, p e14222 (2010)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Teun W J M van Herpen Katarina Cankar Marilise Nogueira Dirk Bosch Harro J Bouwmeester Jules Beekwilder Nicotiana benthamiana as a production platform for artemisinin precursors.
description	<h4>Background</h4>Production of pharmaceuticals in plants provides an alternative for chemical synthesis, fermentation or natural sources. Nicotiana benthamiana is deployed at commercial scale for production of therapeutic proteins. Here the potential of this plant is explored for rapid production of precursors of artemisinin, a sesquiterpenoid compound that is used for malaria treatment.<h4>Methodology/principal findings</h4>Biosynthetic genes leading to artemisinic acid, a precursor of artemisinin, were combined and expressed in N. benthamiana by agro-infiltration. The first committed precursor of artemisinin, amorpha-4,11-diene, was produced upon infiltration of a construct containing amorpha-4,11-diene synthase, accompanied by 3-hydroxy-3-methylglutaryl-CoA reductase and farnesyl diphosphate synthase. Amorpha-4,11-diene was detected both in extracts and in the headspace of the N. benthamiana leaves. When the amorphadiene oxidase CYP71AV1 was co-infiltrated with the amorphadiene-synthesizing construct, the amorpha-4,11-diene levels strongly decreased, suggesting it was oxidized. Surprisingly, no anticipated oxidation products, such as artemisinic acid, were detected upon GC-MS analysis. However, analysis of leaf extracts with a non-targeted metabolomics approach, using LC-QTOF-MS, revealed the presence of another compound, which was identified as artemisinic acid-12-β-diglucoside. This compound accumulated to 39.5 mg x kg(-1) fwt. Apparently the product of the heterologous pathway that was introduced, artemisinic acid, is further metabolized efficiently by glycosyl transferases that are endogenous to N. benthamiana.<h4>Conclusion/significance</h4>This work shows that agroinfiltration of N. bentamiana can be used as a model to study the production of sesquiterpenoid pharmaceutical compounds. The interaction between the ectopically introduced pathway and the endogenous metabolism of the plant is discussed.
format	article
author	Teun W J M van Herpen Katarina Cankar Marilise Nogueira Dirk Bosch Harro J Bouwmeester Jules Beekwilder
author_facet	Teun W J M van Herpen Katarina Cankar Marilise Nogueira Dirk Bosch Harro J Bouwmeester Jules Beekwilder
author_sort	Teun W J M van Herpen
title	Nicotiana benthamiana as a production platform for artemisinin precursors.
title_short	Nicotiana benthamiana as a production platform for artemisinin precursors.
title_full	Nicotiana benthamiana as a production platform for artemisinin precursors.
title_fullStr	Nicotiana benthamiana as a production platform for artemisinin precursors.
title_full_unstemmed	Nicotiana benthamiana as a production platform for artemisinin precursors.
title_sort	nicotiana benthamiana as a production platform for artemisinin precursors.
publisher	Public Library of Science (PLoS)
publishDate	2010
url	https://doaj.org/article/d0864dd6e0444e0b95ce420dd69c3763
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Nicotiana benthamiana as a production platform for artemisinin precursors.

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