High-level production of amorpha-4,11-diene, a precursor of the antimalarial agent artemisinin, in Escherichia coli.

<h4>Background</h4>Artemisinin derivatives are the key active ingredients in Artemisinin combination therapies (ACTs), the most effective therapies available for treatment of malaria. Because the raw material is extracted from plants with long growing seasons, artemisinin is often in sho...

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Autores principales:	Hiroko Tsuruta, Christopher J Paddon, Diana Eng, Jacob R Lenihan, Tizita Horning, Larry C Anthony, Rika Regentin, Jay D Keasling, Neil S Renninger, Jack D Newman
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2009
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/29329a6165654224a54f2846107a793c
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spelling	oai:doaj.org-article:29329a6165654224a54f2846107a793c2021-11-25T06:17:16ZHigh-level production of amorpha-4,11-diene, a precursor of the antimalarial agent artemisinin, in Escherichia coli.1932-620310.1371/journal.pone.0004489https://doaj.org/article/29329a6165654224a54f2846107a793c2009-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/19221601/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Artemisinin derivatives are the key active ingredients in Artemisinin combination therapies (ACTs), the most effective therapies available for treatment of malaria. Because the raw material is extracted from plants with long growing seasons, artemisinin is often in short supply, and fermentation would be an attractive alternative production method to supplement the plant source. Previous work showed that high levels of amorpha-4,11-diene, an artemisinin precursor, can be made in Escherichia coli using a heterologous mevalonate pathway derived from yeast (Saccharomyces cerevisiae), though the reconstructed mevalonate pathway was limited at a particular enzymatic step.<h4>Methodology/ principal findings</h4>By combining improvements in the heterologous mevalonate pathway with a superior fermentation process, commercially relevant titers were achieved in fed-batch fermentations. Yeast genes for HMG-CoA synthase and HMG-CoA reductase (the second and third enzymes in the pathway) were replaced with equivalent genes from Staphylococcus aureus, more than doubling production. Amorpha-4,11-diene titers were further increased by optimizing nitrogen delivery in the fermentation process. Successful cultivation of the improved strain under carbon and nitrogen restriction consistently yielded 90 g/L dry cell weight and an average titer of 27.4 g/L amorpha-4,11-diene.<h4>Conclusions/ significance</h4>Production of >25 g/L amorpha-4,11-diene by fermentation followed by chemical conversion to artemisinin may allow for development of a process to provide an alternative source of artemisinin to be incorporated into ACTs.Hiroko TsurutaChristopher J PaddonDiana EngJacob R LenihanTizita HorningLarry C AnthonyRika RegentinJay D KeaslingNeil S RenningerJack D NewmanPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 4, Iss 2, p e4489 (2009)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Hiroko Tsuruta Christopher J Paddon Diana Eng Jacob R Lenihan Tizita Horning Larry C Anthony Rika Regentin Jay D Keasling Neil S Renninger Jack D Newman High-level production of amorpha-4,11-diene, a precursor of the antimalarial agent artemisinin, in Escherichia coli.
description	<h4>Background</h4>Artemisinin derivatives are the key active ingredients in Artemisinin combination therapies (ACTs), the most effective therapies available for treatment of malaria. Because the raw material is extracted from plants with long growing seasons, artemisinin is often in short supply, and fermentation would be an attractive alternative production method to supplement the plant source. Previous work showed that high levels of amorpha-4,11-diene, an artemisinin precursor, can be made in Escherichia coli using a heterologous mevalonate pathway derived from yeast (Saccharomyces cerevisiae), though the reconstructed mevalonate pathway was limited at a particular enzymatic step.<h4>Methodology/ principal findings</h4>By combining improvements in the heterologous mevalonate pathway with a superior fermentation process, commercially relevant titers were achieved in fed-batch fermentations. Yeast genes for HMG-CoA synthase and HMG-CoA reductase (the second and third enzymes in the pathway) were replaced with equivalent genes from Staphylococcus aureus, more than doubling production. Amorpha-4,11-diene titers were further increased by optimizing nitrogen delivery in the fermentation process. Successful cultivation of the improved strain under carbon and nitrogen restriction consistently yielded 90 g/L dry cell weight and an average titer of 27.4 g/L amorpha-4,11-diene.<h4>Conclusions/ significance</h4>Production of >25 g/L amorpha-4,11-diene by fermentation followed by chemical conversion to artemisinin may allow for development of a process to provide an alternative source of artemisinin to be incorporated into ACTs.
format	article
author	Hiroko Tsuruta Christopher J Paddon Diana Eng Jacob R Lenihan Tizita Horning Larry C Anthony Rika Regentin Jay D Keasling Neil S Renninger Jack D Newman
author_facet	Hiroko Tsuruta Christopher J Paddon Diana Eng Jacob R Lenihan Tizita Horning Larry C Anthony Rika Regentin Jay D Keasling Neil S Renninger Jack D Newman
author_sort	Hiroko Tsuruta
title	High-level production of amorpha-4,11-diene, a precursor of the antimalarial agent artemisinin, in Escherichia coli.
title_short	High-level production of amorpha-4,11-diene, a precursor of the antimalarial agent artemisinin, in Escherichia coli.
title_full	High-level production of amorpha-4,11-diene, a precursor of the antimalarial agent artemisinin, in Escherichia coli.
title_fullStr	High-level production of amorpha-4,11-diene, a precursor of the antimalarial agent artemisinin, in Escherichia coli.
title_full_unstemmed	High-level production of amorpha-4,11-diene, a precursor of the antimalarial agent artemisinin, in Escherichia coli.
title_sort	high-level production of amorpha-4,11-diene, a precursor of the antimalarial agent artemisinin, in escherichia coli.
publisher	Public Library of Science (PLoS)
publishDate	2009
url	https://doaj.org/article/29329a6165654224a54f2846107a793c
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High-level production of amorpha-4,11-diene, a precursor of the antimalarial agent artemisinin, in Escherichia coli.

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