Production of the antimalarial drug precursor artemisinic acid in engineered yeast
Production of the antimalarial drug precursor artemisinic acid in engineered yeast
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DOI：
10.1038/nature04640
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发表时间：
2006-04-13
期刊：
影响因子：
64.8
通讯作者：
Keasling, JD
中科院分区：
文献类型：
--
作者：
Ro, DK;Paradise, EM;Keasling, JD
疟疾是一个全球性的健康问题，威胁着3亿至5亿人，每年造成100多万人死亡（1）。疟疾寄生虫恶性疟原虫（Plasmodium falciparum）的多药耐药菌株的出现阻碍了疾病控制（2，3）。目前正在开发合成抗疟疾药物和疟疾疫苗，但其对疟疾的疗效有待严格的临床试验（4、5）。青蒿素是一种从黄花蒿（菊科;通常称为青蒿）中提取的倍半萜内酯内过氧化物，对多药耐药疟原虫属（Plasmodium spp.）但供应短缺，大多数疟疾患者负担不起（6）。虽然青蒿素的全合成是困难和昂贵的（7），青蒿素或任何衍生物的半合成从微生物来源的青蒿酸，其直接前体，可能是一个成本效益高，环境友好，高质量和可靠的青蒿素来源（8，9）。在这里，我们报告了利用工程化的甲羟戊酸途径、紫穗槐二烯合酶和来自A. annua进行紫穗槐-4，11-二烯到青蒿酸的三步氧化。合成的青蒿酸被运输出来并保留在工程酵母的外部，这意味着可以使用简单且廉价的纯化过程来获得所需的产品。尽管工程酵母已经能够以比A.要将青蒿酸产量提高到足以将青蒿素综合疗法减少到大大低于其当前价格的水平，就需要进行《青蒿素年度方案》、优化产量和扩大工业规模。
Malaria is a global health problem that threatens 300-500 million people and kills more than one million people annually(1). Disease control is hampered by the occurrence of multi-drug-resistant strains of the malaria parasite Plasmodium falciparum(2,3). Synthetic antimalarial drugs and malarial vaccines are currently being developed, but their efficacy against malaria awaits rigorous clinical testing(4,5). Artemisinin, a sesquiterpene lactone endoperoxide extracted from Artemisia annua L (family Asteraceae; commonly known as sweet wormwood), is highly effective against multi-drug-resistant Plasmodium spp., but is in short supply and unaffordable to most malaria sufferers(6). Although total synthesis of artemisinin is difficult and costly(7), the semi-synthesis of artemisinin or any derivative from microbially sourced artemisinic acid, its immediate precursor, could be a cost-effective, environmentally friendly, high-quality and reliable source of artemisinin(8,9). Here we report the engineering of Saccharomyces cerevisiae to produce high titres (up to 100 mg l(-1)) of artemisinic acid using an engineered mevalonate pathway, amorphadiene synthase, and a novel cytochrome P450 monooxygenase (CYP71AV1) from A. annua that performs a three-step oxidation of amorpha-4,11-diene to artemisinic acid. The synthesized artemisinic acid is transported out and retained on the outside of the engineered yeast, meaning that a simple and inexpensive purification process can be used to obtain the desired product. Although the engineered yeast is already capable of producing artemisinic acid at a significantly higher specific productivity than A. annua, yield optimization and industrial scale-up will be required to raise artemisinic acid production to a level high enough to reduce artemisinin combination therapies to significantly below their current prices.