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Metabolic engineering for the production of clinically important molecules: Omega-3 fatty acids, artemisinin, and taxol
Article first published online: 21 OCT 2011
DOI: 10.1002/biot.201100289
Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Issue

Biotechnology Journal
Special Issue: Biotech Methods and Advances
Volume 7, Issue 1, pages 20–33, January 2012
Additional Information
How to Cite
Ye, V. M. and Bhatia, S. K. (2012), Metabolic engineering for the production of clinically important molecules: Omega-3 fatty acids, artemisinin, and taxol. Biotechnology Journal, 7: 20–33. doi: 10.1002/biot.201100289
Publication History
- Issue published online: 9 JAN 2012
- Article first published online: 21 OCT 2011
- Manuscript Accepted: 2 SEP 2011
- Manuscript Revised: 28 AUG 2011
- Manuscript Received: 30 JUL 2011
REFERENCES
- 1, Manufacturing molecules through metabolic engineering. Science 2010, 330, 1355–1358.
- 2, Tissue engineering for clinical applications. Biotechnol. J. 2010, 5, 1309–1323.Direct Link:
- 3, , , et al., Metabolic engineering of microorganisms: General strategies and drug production. Drug Discov. Today 2009, 14, 78–88.
- 4, , , , Differences between dietary supplement and prescription drug omega-3 fatty acid formulations: A legislative and regulatory perspective. J. Am. Coll. Nutr. 2008, 27, 659–666.
- 5, , , et al., Effects of eicosapentaenoic acid on major coronary events in hypercholesterolaemic patients (JELIS): A randomised open-label, blinded endpoint analysis. Lancet 2007, 369, 1090–1098.
- 6, , , et al., multiple risk factors: Sub-analysis of primary prevention cases from the Japan EPA Lipid Intervention Study (JELIS). Atherosclerosis 2008, 200, 135–140.
- 7GISSI-HF Investigators, Effect of n-3 polyunsaturated fatty acids in patients with chronic heart failure (the GISSI-HF trial): A randomised, double-blind, placebo-controlled trial. Lancet 2008, 372, 1223–1230.
- 8, , , Dietary reference intakes for DHA and EPA. Prostaglandins Leukot. Essent. Fatty Acids 2009, 81, 99–104.
- 9, , , et al., Reduction in the recurrence of stroke by eicosapentaenoic acid for hypercholesterolemic patients: Subanalysis of the JELIS trial. Stroke 2008, 39, 2052–2058.
- 10, , , et al., Plasma n-3 fatty acids and the risk of cognitive decline in older adults: The Atherosclerosis risk in communities study. Am. J. Clin. Nutr. 2007, 85, 1103–1111.
- 11, N-3 Polyunsaturated fatty acids, inflammation, and inflammatory diseases. Am. J. Clin. Nutr. 2006, 83, 1505S–1519S.
- 12
- 13
- 14Zhu, Q., Xue, Z., Yadav, N., Damude, H. et al., Metabolic engineering of an oleaginous yeast for the production of omega-3 fatty acids, in: Cohen Z., Ratledge C. (Eds.), Single Cell Oil, 2nd Edn., ACOS Press, Urbana, IL 2010, pp. 51–73.
- 15, , , et al., The same rat Delta6-desaturase not only acts on 18- but also on 24-carbon fatty acids in very-long-chain polyunsaturated fatty acid biosynthesis. Biochem. J. 2002, 364, 49–55.
- 16, The roles of anabolic and catabolic reactions in the synthesis and recycling of polyunsaturated fatty acids. Prostaglandins Leukot. Essent. Fatty Acids 2002, 67, 79–83.
- 17, , , , Polyunsaturated fatty acids: Biochemical, nutritional and epigenetic properties. J. Am. Coll. Nutr. 2004, 23, 281–302.
- 18, , Conversion of alpha-linolenic acid to longer-chain polyunsaturated fatty acids in human adults. Reprod. Nutr. Dev. 2005, 45, 581–597.
- 19, , et al., n-3 fatty acids from fish or fish-oil supplements, but not alpha-linolenic acid, benefit cardiovascular disease outcomes in primary- and secondary-prevention studies: A systematic review. Am J. Clin. Nutr. 2006, 84, 5–17.
- 20, , , , Contaminants in fish tissue from US lakes and reservoirs: A national probabilistic study. Environ. Monit. Assess. 2009, 150, 3–19.
- 21, , Levels of PCDD/PCDFs and PCBs in edible marine species and human intake: A literature review. Environ. Int. 2007, 33, 397–405.
- 22, , Lipid production by Yarrowia lipolytica growing on industrial glycerol in a single-stage continuous culture. Bioresour. Technol. 2002, 82, 43–49.
- 23, , , et al., Single cell oil production by Yarrowia lipolytica growing on an industrial derivative of animal fat in batch cultures. Appl. Microbiol. Biotechnol. 2002, 58, 308–312.
- 24, , , , , Insertional mutagenesis in the n-alkane-assimilating yeast Yarrowia lipolytica: Generation of tagged mutations in genes involved in hydrophobic substrate utilization. J. Bacteriol. 2001, 183, 5102–5109.
- 25, , , , Identification of bifunctional Delta 12/omega 3 fatty acid desaturases for improving the ratio of omega 3 to omega 6 fatty acids in microbes and plants. Proc. Natl. Acad. Sci. USA 2006, 103, 9446–9451.
- 26Damude, H. G., Gillies, P. J., Macool, D. J., Picataggio, S. K. et al., High eicosapentaenoic acid producing strains of Yarrowia lipolytica. US Patent 07932077, 2011.
- 27, , , , Safety assessment of EPA-rich triglyceride oil produced from yeast: Genotoxicity and 28-day oral toxicity in rats. Regul. Toxicol. Pharmacol. 2010, 59, 53–63.
- 28, , , et al., Safety assessment of EPA-rich oil produced from yeast: Results of a 90-day subchronic toxicity study. Regul. Toxicol. Pharmacol. 2010, 58, 490–500.
- 29, , , Metabolism of stearidonic acid in human subjects: Comparison with the metabolism of other n-3 fatty acids. Am. J. Clin. Nutr. 2003, 77, 1140–1145.
- 30, , , et al., The synthesis and accumulation of stearidonic acid in transgenic plants: A novel source of 'heart-healthy' omega-3 fatty acids. Plant Biotechnol. J. 2009, 7, 704–716.Direct Link:
- 31, Dietary stearidonic acid is a long chain (n-3) polyunsaturated fatty acid with potential health benefits. J. Nutr. 2009, 139, 5–10.
- 32Monsanto Company, Stearidonic (SDA) Omega-3 Soybean Oil GRAS Notice 2009. (http://www.accessdata.fda.gov/scripts/fcn/gras_notices/grn000283.pdf).
- 33, , , , Dietary intake of stearidonic acid-enriched soybean oil increases the omega-3 index: Randomized, double-blind clinical study of efficacy and safety. Am. J. Clin. Nutr. 2010, 92, 766–75.
- 34, , et al., Stearidonic acid-enriched soybean oil increased the omega-3 index, an emerging cardiovascular risk marker. Lipids 2008, 43, 805–811.
- 35US Food and Drug Administration, Agency Response Letter GRAS Notice No. GRN 000283 2009. (http://www.fda.gov/Food/FoodIngredientsPackaging/GenerallyRecognizedasSafeGRAS/GRASListings/ucm185688.htm).
- 36, , Enhancing plant seed oils for human nutrition. Plant Physiol. 2008, 147, 962–968.
- 37, , Knowledge of the proposed chemical mechanism of action and cytochrome p450 metabolism of antimalarial trioxanes like artemisinin allows rational design of new antimalarial peroxides. Acc. Chem. Res. 2004, 37, 397–404.
- 38, , , et al., Artemisinin activity against Plasmodium falciparum requires hemoglobin uptake and digestion. Proc. Natl. Acad. Sci. USA 2011, 108, 11405–11410.
- 39, , , , Artemisinins act through at least two targets in a yeast model. FEMS Yeast Res. 2011, 11, 233–7.Direct Link:
- 40, , , et al., Artemisinin directly targets malarial mitochondria through its specific mitochondrial activation. PLoS One 2010, 5, e9582.
- 41, , Anticancer activities of artemisinin and its bioactive derivatives. Expert. Rev. Mol. Med. 2009, 11, e32.
- 42World Health Organization, World Malaria Report 2008, Geneva: WHO Press 2008.
- 43, Qinghaosu (Artemisinin): The Price of Success. Science 2008, 18, 330–334.
- 44Artepal, J. P., Artemisinin Market: Quantities and pricing. Artemisinin Conference 2010, Antananarivo, Madagascar.
- 45, The non-mevalonate pathway of isoprenoid precursor biosynthesis. J. Biol. Chem. 2007, 282, 21573–21577.
- 46, , , , Isoprenoid biosynthesis: The evolution of two ancient and distinct pathways across genomes. Proc. Natl. Acad. Sci. USA 2000, 97, 13172–13177.
- 47, , , et al., Amorpha-4,11-diene synthase catalyses the first probable step in artemisinin biosynthesis. Phytochemistry 1999, 52, 843–854.
- 48, , , et al., Production of the antimalarial drug precursor artemisinic acid in engineered yeast. Nature 2006, 440, 940–943.
- 49, , , et al., Artemisia annua L. (Asteraceae) trichome-specific cDNAs reveal CYP71AV1, a cytochrome P450 with a key role in the biosynthesis of the antimalarial sesquiterpene lactone artemisinin. FEBS Lett. 2006, 580, 1411–1416.
- 50, , , et al., The molecular cloning of artemisinic aldehyde Delta11(13) reductase and its role in glandular trichome-dependent biosynthesis of artemisinin in Artemisia annua. J. Biol. Chem. 2008, 283, 21501–21508.
- 51, , , , Molecular cloning of an aldehyde dehydrogenase implicated in artemisinin biosynthesis in Artemisia annua. Botany 2009, 87, 635–642.
- 52, , , , , Engineering a mevalonate pathway in Escherichia coli for production of terpenoids. Nat. Biotechnol. 2003, 21, 796–802.
- 53, , , , Balancing a heterologous mevalonate pathway for improved isoprenoid production in Escherichia coli. Metab. Eng. 2007, 9, 193–207.
- 54, , , et al., High-level production of amorpha-4,11-diene, a precursor of the antimalarial agent artemisinin, in Escherichia coli. PLoS One 2009, 4, e4489.
- 55, , , Effects of overproduction of the catalytic domain of 3-hydroxy-3-methylglutaryl coenzyme A reductase on squalene synthesis in Saccharomyces cerevisiae. Appl. Environ. Microbiol. 1997, 63, 3341–3344.
- 56, , Upc2p and Ecm22p, dual regulators of sterol biosynthesis in Saccharomyces cerevisiae. Mol. Cell. Biol. 2001, 21, 6395–6405.
- 57, , , et al., , Identification of intermediates and enzymes involved in the early steps of artemisinin biosynthesis in Artemisia annua. Planta Med. 2005, 71, 40–47.
- 58Farret, H., Semi-Synthetic Artemisinin Project. Artemisinin Conference 2010, Antananarivo, Madagascar.
- 59Nguyen, T. H., Semi-Synthetic Project One World Health. Artemisinin Conference 2010, Antananarivo, Madagascar.
- 60, , , , Overexpression of the HMG-CoA reductase gene leads to enhanced artemisinin biosynthesis in transgenic Artemisia annua plants. Planta Med. 2009, 75, 1453–1458.
- 61, , Over-expression of HMG-CoA reductase and amorpha-4,11-diene synthase genes in Artemisia annua L. and its influence on artemisinin content. Plant Cell Rep. 2011. 30, 1919–1928.
- 62
- 63, , , et al., Development of transgenic Artemisia annua (Chinese wormwood) plants with an enhanced content of artemisinin, an effective anti-malarial drug, by hairpin-RNA-mediated gene silencing Biotechnol. Appl. Biochem. 2009, 52, 199–207.Direct Link:
- 64, , , , The production of artemisinin precursors in tobacco. Plant Biotechnol. J. 2011, 9, 445–454.Direct Link:
- 65, , , et al., Nicotiana benthamiana as a production platform for artemisinin precursors. PLoS One 2010, 5, e14222.
- 66, , , et al., Genetic engineering of terpenoid metabolism attracts, bodyguards to Arabidopsis. Science 2005, 309, 2070–2072.
- 67, , , et al., Analysis of the aphthovirus 2A/2B polyprotein 'cleavage' mechanism indicates not a proteolytic reaction, but a novel translational effect: A putative ribosomal 'skip'. J. Gen. Virol. 2001, 82, 1013–1025.
- 68, , , et al., Building a golden triangle for the production and use of artemisinin derivatives against falciparum malaria in Africa. Afr. J. Biotechnol. 2008, 7, 4884–4896.
- 69Dafra Pharma, Chicory Project (http://www.dafra.be/content/chicory-project).
- 70, , , , Paclitaxel in cancer treatment. Semin Oncol. 1992, 19, 646–662.
- 71, , , Promotion of microtubule assembly in vitro by taxol. Nature 1979, 277, 665–667.
- 72, , , et al., Medicinal chemistry of paclitaxel and its analogues. Curr. Med. Chem. 2009, 16, 3966–3985.
- 73, , , Paclitaxel inhibits arterial smooth muscle cell proliferation and migration in vitro and in vivo using local drug delivery. Circulation 1997, 96, 636–645.
- 74, , , A critical appraisal of the safety and efficacy of drug-eluting stents. Clin. Pharmacol. Ther. 2009, 85, 474–480.
- 75, , , et al., Paclitaxel and immune system. Eur. J. Pharm. Sci. 2009, 38, 283–290.
- 76, , , Paclitaxel-loaded polymeric micelles modified with MCF-7 cell-specific phage protein: Enhanced binding to target cancer cells and increased cytotoxicity. Mol. Pharm. 2010, 7, 1007–1014.
- 77, , , , Palmitoyl ascorbate-modified liposomes as nanoparticle platform for ascorbate-mediated cytotoxicity and paclitaxel co-delivery. Eur. J. Pharm. Biopharm. 2010, 75, 321–326.
- 78, , Camptothecin and taxol: Discovery to clinic-thirteenth Bruce F. Cain Memorial Award Lecture. Cancer Res. 1995, 55, 753–760.
- 79, , , et al., Plant antitumor agents. VI. The isolation and structure of taxol, a novel antileukemic and antitumor agent from Taxus brevifolia. J. Am. Chem. Soc. 1971, 93, 2325–2327.
- 80Goodman, J., Walsh V., The story of taxol: Nature and politics in the pursuit of an anti-cancer drug. Cambridge University Press, Cambridge 2001, 193–249.
- 81, , , et al., First total synthesis of taxol. 2. completion of the C and D rings. J. Am. Chem. Soc. 1994, 116, 1599–1600.
- 82, , , et al., First total synthesis of taxol. 1. Functionalization of the B ring. J. Am. Chem. Soc. 1994, 116, 1597–1598.
- 83, , , et al., Total synthesis of taxol. Nature 1994, 367, 630–634.
- 84Holton, R. A., Biediger, R. J., Boatman, P. D., Semisynthesis of taxol and taxotere. in: Suffness M. (Ed.), Taxol Science and Applications. CRC Press, Boca Raton 1995, pp. 97–121.
- 85Phyton Biotech. 2010. (http://www.phytonbiotech.com/index.htm).
- 86, , Taxol biosynthetic genes. Phytochemistry 2001, 58, 1–7.
- 87
- 88, , , , Cytochrome p450 taxadiene 5alpha-hydroxylase, a mechanistically unusual monooxygenase catalyzing the first oxygenation step of taxol biosynthesis. Chem Biol. 2004, 11, 379–87.
- 89, , , , Specificity of the N-benzoyl transferase responsible for the last step of Taxol biosynthesis. Arch. Biochem. Biophys. 2008, 477, 384–389.
- 90, , , et al., Genetic engineering of taxol biosynthetic genes in Saccharomyces cerevisiae. Biotechnol. Bioeng. 2006, 93, 212–224.Direct Link:
- 91, , , Metabolic engineering of taxadiene biosynthesis in yeast as a first step towards Taxol (Paclitaxel) production. Metab. Eng. 2008, 10, 201–206.
- 92, , , et al., Isoprenoid pathway optimization for Taxol precursor overproduction in Escherichia coli. Science 2010, 330, 70–74.
- 93, , Production of isoprenoid pharmaceuticals by engineered microbes. Nat. Chem. Biol. 2006, 2, 674–681.
- 94, , Engineering of artificial plant cytochrome P450 enzymes for synthesis of isoflavones by Escherichia coli. Appl. Environ. Microbiol. 2007, 73, 7246–7251.

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