Publication Date:
2010-10-12
Description:
Taxol (paclitaxel) is a potent anticancer drug first isolated from the Taxus brevifolia Pacific yew tree. Currently, cost-efficient production of Taxol and its analogs remains limited. Here, we report a multivariate-modular approach to metabolic-pathway engineering that succeeded in increasing titers of taxadiene--the first committed Taxol intermediate--approximately 1 gram per liter (~15,000-fold) in an engineered Escherichia coli strain. Our approach partitioned the taxadiene metabolic pathway into two modules: a native upstream methylerythritol-phosphate (MEP) pathway forming isopentenyl pyrophosphate and a heterologous downstream terpenoid-forming pathway. Systematic multivariate search identified conditions that optimally balance the two pathway modules so as to maximize the taxadiene production with minimal accumulation of indole, which is an inhibitory compound found here. We also engineered the next step in Taxol biosynthesis, a P450-mediated 5alpha-oxidation of taxadiene to taxadien-5alpha-ol. More broadly, the modular pathway engineering approach helped to unlock the potential of the MEP pathway for the engineered production of terpenoid natural products.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3034138/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉 〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3034138/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Ajikumar, Parayil Kumaran -- Xiao, Wen-Hai -- Tyo, Keith E J -- Wang, Yong -- Simeon, Fritz -- Leonard, Effendi -- Mucha, Oliver -- Phon, Too Heng -- Pfeifer, Blaine -- Stephanopoulos, Gregory -- 1-R01-GM085323-01A1/GM/NIGMS NIH HHS/ -- R01 GM085323/GM/NIGMS NIH HHS/ -- R01 GM085323-02/GM/NIGMS NIH HHS/ -- New York, N.Y. -- Science. 2010 Oct 1;330(6000):70-4. doi: 10.1126/science.1191652.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Chemical Engineering, Massachusetts Institute of Technology (MIT), Cambridge, MA 02139, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/20929806" target="_blank"〉PubMed〈/a〉
Keywords:
Alkenes/*metabolism
;
Bioreactors
;
Cytochrome P-450 Enzyme System/genetics/metabolism
;
Diterpenes/*metabolism
;
Erythritol/analogs & derivatives/metabolism
;
Escherichia coli K12/enzymology/genetics/*metabolism
;
Farnesyltranstransferase/genetics/metabolism
;
Fermentation
;
*Genetic Engineering
;
Hemiterpenes/metabolism
;
Indoles/metabolism
;
Isomerases/genetics/metabolism
;
Metabolic Networks and Pathways/genetics
;
Metabolomics
;
NADPH-Ferrihemoprotein Reductase/genetics/metabolism
;
Organophosphorus Compounds/metabolism
;
Oxidation-Reduction
;
Paclitaxel/*biosynthesis
;
Recombinant Fusion Proteins/metabolism
;
Sugar Phosphates/metabolism
;
Taxoids/metabolism
;
Taxus/enzymology
;
Terpenes/metabolism
Print ISSN:
0036-8075
Electronic ISSN:
1095-9203
Topics:
Biology
,
Chemistry and Pharmacology
,
Computer Science
,
Medicine
,
Natural Sciences in General
,
Physics
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