Artemisinin (AN), a sesquiterpene lactone, and its derivatives are proven, effective antimalarial therapeutics that also show anticancer activity. However, the complex compound is still best produced in plants; and the drug is in short supply. AN can be produced in transformed root cultures of Artemisa annua L.; we have considerable experience with controlling production. Since to our knowledge it has not yet been established which of the two arms of the terpenoid biosynthetic pathway are operational in the biosynthesis of artemisinin, we hypothesize that AN is produced from a pool of IPP that originates from both the mevalonate and the non-mevalonate pathway, as shown for sesquiterpenes of chamomile, a close relative of Artemisia annua L. Since the sesquiterpene cyclase, ADS, leads to AN and has recently been, we hypothesize that this gene is also present and responsive in our transformed roots that are known producers of artemisinin. Objectives: to be accomplished over 2.5 years (% effort): 1. To do C13 labeling experiments to determine the fate of carbon in artemisinin biosynthesis so that we might know whether one, the other, or both arms of the terpenoid biosynthetic pathway are playing a role in its production. (65%) 2. To produce a molecular probe for the specific sesquiterpene cyclase (SQC) gene, amorpha-4,11-diene synthase (ADS), that leads to artemisinin in order to study any future ADS expression in hairy roots if roots ire ever to be considered as a production system for artemisinin. (15%) 3. To demonstrate that expression of the ADS probe correlates with artemisinin production using conditions where production is known to be either very high or very low. (20%) The long-term goal of our work is to improve fundamental understanding of the regulatory control of terpene production in hairy roots so that we can demonstrate their utility as a production system for important secondary metabolites. This study will enhance our understanding of the biosynthesis of AN in plants and further our ability to increase production of artemisinin while improving overall understanding of terpenoid biosynthesis.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Academic Research Enhancement Awards (AREA) (R15)
Project #
1R15GM069562-01
Application #
6702193
Study Section
Bio-Organic and Natural Products Chemistry Study Section (BNP)
Program Officer
Jones, Warren
Project Start
2004-04-01
Project End
2007-03-31
Budget Start
2004-04-01
Budget End
2007-03-31
Support Year
1
Fiscal Year
2004
Total Cost
$192,550
Indirect Cost
Name
Worcester Polytechnic Institute
Department
Biology
Type
Schools of Engineering
DUNS #
041508581
City
Worcester
State
MA
Country
United States
Zip Code
01609
Weathers, Pamela J; Elfawal, Mostafa A; Towler, Melissa J et al. (2014) Pharmacokinetics of artemisinin delivered by oral consumption of Artemisia annua dried leaves in healthy vs. Plasmodium chabaudi-infected mice. J Ethnopharmacol 153:732-6
Weathers, Pamela J; Jordan, Nikole J; Lasin, Praphapan et al. (2014) Simulated digestion of dried leaves of Artemisia annua consumed as a treatment (pACT) for malaria. J Ethnopharmacol 151:858-63
Nguyen, Khanhvan T; Towler, Melissa J; Weathers, Pamela J (2013) The effect of roots and media constituents on trichomes and artemisinin production in Artemisia annua L. Plant Cell Rep 32:207-18
Weathers, Pamela J; Towler, Melissa J (2012) The flavonoids casticin and artemetin are poorly extracted and are unstable in an Artemisia annua tea infusion. Planta Med 78:1024-6
Weathers, Pamela J; Arsenault, Patrick R; Covello, Patrick S et al. (2011) Artemisinin production in Artemisia annua: studies in planta and results of a novel delivery method for treating malaria and other neglected diseases. Phytochem Rev 10:173-183
Nguyen, Khanhvan T; Arsenault, Patrick R; Weathers, Pamela J (2011) Trichomes + roots + ROS = artemisinin: regulating artemisinin biosynthesis in Artemisia annua L. In Vitro Cell Dev Biol Plant 47:329-338
Arsenault, Patrick R; Vail, Daniel; Wobbe, Kristin K et al. (2010) Reproductive development modulates gene expression and metabolite levels with possible feedback inhibition of artemisinin in Artemisia annua. Plant Physiol 154:958-68
Sivakumar, Ganapathy; Liu, Chunzhao; Towler, Melissa J et al. (2010) Biomass production of hairy roots of Artemisia annua and Arachis hypogaea in a scaled-up mist bioreactor. Biotechnol Bioeng 107:802-13
Arsenault, Patrick R; Vail, Daniel R; Wobbe, Kristin K et al. (2010) Effect of sugars on artemisinin production in Artemisia annua L.: transcription and metabolite measurements. Molecules 15:2302-18
Mannan, Abdul; Liu, Chunzhao; Arsenault, Patrick R et al. (2010) DMSO triggers the generation of ROS leading to an increase in artemisinin and dihydroartemisinic acid in Artemisia annua shoot cultures. Plant Cell Rep 29:143-52

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