izpis_h1_title_alt

Nicotiana benthamiana as a production platform for artemisinin precursors
Harpen, Teun W. J. M. van (Author), Cankar, Katarina (Author), Nogueira, Marilise (Author), Bosch, Dirk (Author), Bouwmeester, Harro (Author), Beekwilder, Jules (Author)

URLURL - Presentation file, Visit http://dx.doi.org/10.1371/journal.pone.0014222 New window

Abstract
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.

Language:English
Keywords:pharmaceuticals, therapeutic proteins, malaria, biosynthesis
Work type:Not categorized (r6)
Tipology:1.01 - Original Scientific Article
Organization:BF - Biotechnical Faculty
Year:2010
Number of pages:str. 1-11
Numbering:Vol. 5, no. 12
UDC:581
ISSN on article:1932-6203
DOI:10.1371/journal.pone.0014222 Link is opened in a new window
COBISS.SI-ID:2323535 Link is opened in a new window
Views:524
Downloads:161
Metadata:XML RDF-CHPDL DC-XML DC-RDF
 
Average score:(0 votes)
Your score:Voting is allowed only to logged in users.
:
Share: Bookmark and Share

Record is a part of a journal

Title:PloS one
Publisher:Public Library of Science
COBISS.SI-ID:2005896 New window

Similar documents

Similar works from RUL:
Similar works from other Slovenian collections:

Comments

Leave comment

You have to log in to leave a comment.

Comments (0)
0 - 0 / 0
 
There are no comments!

Back