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Nicotiana benthamiana as a production platform for artemisinin precursorsHarpen, Teun W. J. M. van ...BACKGROUND: Production of pharmaceuticals in plants provides an alternative for chemical synthesis, fermentation or natural sources. Nicotiana benthamianais deployed at commercial scale for ... production of therapeutic proteins. Here the potential of this plant is explored for rapid production ofprecursors of artemisinin, a sesquiterpenoid compound that is used for malaria treatment. METHODOLOGY/PRINCIPAL FINDINGS: 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 constructcontaining 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 ofanother compound, which was identified as artemisinic acid-12-beta-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. CONCLUSION/SIGNIFICANCE: 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.Source: PloS one [Elektronski vir]. - ISSN 1932-6203 (Vol. 5, no. 12, 2010, str. 1-11)Type of material - article, component part ; adult, seriousPublish date - 2010Language - englishCOBISS.SI-ID - 2323535
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Database name | Field | Year |
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Links to authors' personal bibliographies | Links to information on researchers in the SICRIS system |
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Harpen, Teun W. J. M. van | |
Cankar, Katarina | 23607 |
Nogueira, Marilise | |
Bosch, Dirk | |
Bouwmeester, Harro | |
Beekwilder, Jules |
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