Document Detail


Production of amorphadiene in yeast, and its conversion to dihydroartemisinic acid, precursor to the antimalarial agent artemisinin.
MedLine Citation:
PMID:  22247290     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Malaria, caused by Plasmodium sp, results in almost one million deaths and over 200 million new infections annually. The World Health Organization has recommended that artemisinin-based combination therapies be used for treatment of malaria. Artemisinin is a sesquiterpene lactone isolated from the plant Artemisia annua. However, the supply and price of artemisinin fluctuate greatly, and an alternative production method would be valuable to increase availability. We describe progress toward the goal of developing a supply of semisynthetic artemisinin based on production of the artemisinin precursor amorpha-4,11-diene by fermentation from engineered Saccharomyces cerevisiae, and its chemical conversion to dihydroartemisinic acid, which can be subsequently converted to artemisinin. Previous efforts to produce artemisinin precursors used S. cerevisiae S288C overexpressing selected genes of the mevalonate pathway [Ro et al. (2006) Nature 440:940-943]. We have now overexpressed every enzyme of the mevalonate pathway to ERG20 in S. cerevisiae CEN.PK2, and compared production to CEN.PK2 engineered identically to the previously engineered S288C strain. Overexpressing every enzyme of the mevalonate pathway doubled artemisinic acid production, however, amorpha-4,11-diene production was 10-fold higher than artemisinic acid. We therefore focused on amorpha-4,11-diene production. Development of fermentation processes for the reengineered CEN.PK2 amorpha-4,11-diene strain led to production of > 40 g/L product. A chemical process was developed to convert amorpha-4,11-diene to dihydroartemisinic acid, which could subsequently be converted to artemisinin. The strains and procedures described represent a complete process for production of semisynthetic artemisinin.
Authors:
Patrick J Westfall; Douglas J Pitera; Jacob R Lenihan; Diana Eng; Frank X Woolard; Rika Regentin; Tizita Horning; Hiroko Tsuruta; David J Melis; Andrew Owens; Scott Fickes; Don Diola; Kirsten R Benjamin; Jay D Keasling; Michael D Leavell; Derek J McPhee; Neil S Renninger; Jack D Newman; Chris J Paddon
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-01-12
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  109     ISSN:  1091-6490     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  2012 Jan 
Date Detail:
Created Date:  2012-02-06     Completed Date:  2012-03-21     Revised Date:  2013-06-26    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  E111-8     Citation Subset:  IM    
Affiliation:
Amyris, Inc., 5885 Hollis Street, Suite 100, Emeryville, CA 94608, USA.
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MeSH Terms
Descriptor/Qualifier:
Antimalarials / chemistry,  metabolism*
Artemisinins / chemistry,  metabolism*
Batch Cell Culture Techniques
Codon / genetics
Ethanol / metabolism
Fermentation
Galactose / metabolism
Genes, Fungal / genetics
Genotype
Glucose / metabolism
Saccharomyces cerevisiae / genetics,  metabolism*
Sesquiterpenes / chemistry,  metabolism*
Chemical
Reg. No./Substance:
0/Antimalarials; 0/Artemisinins; 0/Codon; 0/Sesquiterpenes; 0/amorpha-4,11-diene; 0/dihydroartemisinic acid; 26566-61-0/Galactose; 50-99-7/Glucose; 64-17-5/Ethanol; 9RMU91N5K2/artemisinine
Comments/Corrections

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