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Compartmentalized Metabolic Engineering for Artemisinin biosynthesis and Effective Malaria Treatment by Oral Delivery of Plant Cells

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Molecular Plant 2016年第11期9卷 1464-1477页

作者： Karan Malhotra Mayavan Subramaniyan Khushboo Rawat Md. Kalamuddin M. Irfan Qureshi Pawan Malhotra Asif Mohmmed Katrina Cornish Henry Daniell Shashi Kumar1 Metabolic Engineering Group Malaria Research Group International Centre for Genetic Engineering and Biotechnology Aruna Asaf Ali Marg New Delhi 110067 India Department of Biotechnology Jamia Millia Islamia New Delhi 110025 India Department of Horticulture and Crop Science The Ohio State University Wooster OH 44691 USA Department of Biochemistry School of Dental Medicine University of Pennsylvania Philadelphia PA 19104 USA

Artemisinin is highly effective against drug-resistant malarial parasites, which affects nearly half of the global population and kills 〉500 000 people each year. The primary cost of artemisinin is the very expensive process used to extract and purify the drug from Artemisia annua. Elimination of this apparently unnecessary step will make this potent antimalarial drug affordable to the global population living in endemic regions. Here we reported the oral delivery of a non-protein drug artemisinin biosynthesized （~0.8 mg/g dry weight） at clinically meaningful levels in tobacco by engineering two metabolic pathways targeted to three different cellular compartments （chloroplast, nucleus, and mitochondria）. The doubly transgenic lines showed a three-fold enhancement of isopentenyl pyrophosphate, and targeting AACPR, DBR2, and CYP71AV1 to chloroplasts resulted in higher expression and an efficient photo-oxidation of di- hydroartemisinic acid to artemisinin. Partially purified extracts from the leaves of transgenic tobacco plants inhibited in vitro growth progression of Plasmodium falciparum-infected red blood cells. Oral feeding of whole intact plant cells bioencapsulating the artemisinin reduced the parasitemia levels in challenged mice in comparison with commercial drug. Such novel synergistic approaches should facilitate low-cost production and delivery of artemisinin and other drugs through metabolic engineering of edible plants.

关键词： bioencapsulation oral delivery of plant material dihydroartemisinic acid isopentenyl pyrophosphate,plant transformation drug biosynthesis

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