Genetic engineering of artemisinin biosynthesis: prospects to improve its production

• Autorzy: Abdin N.Z. , Alam P.
• Języki publikacji: EN

Abstrakty

EN

Artemisinin, a potent antimalarial natural compound, is obtained from aerial parts of Artemisia annua L. plants. The demand (101–119 MT) for artemisinin is exponentially increasing every year because of increased incidence of drug-resistant malaria throughout the world, especially in Africa and Asia. However, the presence of low concentrations (0.01–1.1 %) of the compound in A. annua L. plants poses a major constraint in the commercialization of artemisinin-based combination therapies (ACTs) recommended by WHO for the treatment of multidrug- resistant and cerebral malaria. Further, the improvement in the yield of artemisinin through conventional breeding, in vitro culture, cell suspension culture and total organic synthesis still poses a challenge. However, possibilities are there to enhance the artemisinin biosynthesis either by overexpression of the genes encoding enzymes associated with the rate-limiting steps of the mevalonate and artemisinin biosynthetic pathways or by the suppression of genes encoding enzymes of other competing pathways. Based on the current understanding of the pathway and cloning of the related genes, efforts have been made for the past few years to increase the artemisinin content in A. annua L., Cichorium intybus L. and microbes through metabolic engineering. In the present review, we have discussed the metabolic engineering strategies in both plant and microbial systems for artemisinin accumulation in bioengineered hosts.

• Wydawca: -
• Czasopismo: Acta Physiologiae Plantarum
• Rocznik: 2015
• Tom: 37
• Numer: 02
• Opis fizyczny: Article: 33 [12 p.], fig.,ref.

Twórcy

autor

Abdin N.Z.

• Centre for Transgenic Plant Development, Department of Biotechnology, Jamia Hamdard, New Delhi 110062, India

autor

Alam P.

• Department of Biosciences, Jamia Millia Islamia, New Delhi 110025, India

Bibliografia

• Abdin MZ, Israr M, Rehman RU, Jain SK (2003) Artemisinin, a novel antimalarial drug: biochemical and molecular approaches for enhanced production. Planta Med 69:289–299
• Ahlawat S, Saxena P, Alam P, Mauji Ram, Nafis T, Mohd A, Abdin MZ (2012) Influence of Agrobacterium rhizogenes on induction of hairy roots for enhanced production of artemisinin in Artemisia annua L. plants. Afr J Biotecnol 1:8684–8691
• Akhila A, Kumkum R, Thakur RS (1999) Biosynthesis of artemisinic acid in Artemisia annua. Phytochemistry 29:2129–2132
• Akiyoshi DE, Morris RO, Hinz R, Mischke BS, Kosuge T, Garfinkel DJ, Gordon MP, Nester EW (1983) Cytokinin/auxin balance in crown gall tumors is regulated by specific loci in the T-DNA. Proc Natl Acad Sci USA 80(2):407–411
• Akiyoshi DE, Klee H, Amasino RM, Nester EW, Gordon MP (1984) T-DNA of Agrobacterium tumefaciens encodes an enzyme of cytokinin biosynthesis. Proc Natl Acad Sci USA 81:5994–5998
• Alam, P, Kamaluddin, Khan MA, Mohammad A, Khan R, Abdin MZ (2014) Enhanced artemisinin accumulation and metabolic profiling of transgenic Artemisia annua L. plants over-expressing by rate limiting enzymes from isoprenoid pathway, J Plant Inter. doi:10.1080/17429145.2014.893030
• Alam P, Abdin MZ (2011) 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 10:1919–1928
• Alam P, Kiran U, Ahmad MM, Kamaluddin Khan MA, Jhanwar S, Abdin MZ (2010) Isolation, characterization and structuralfeatures of amorpha—4,11-diene synthase (ADS3963) from Artemisia annua L. Bioinformation 4:421–429
• Allen RS, Millgate AG, Chitty JA, Thisleton J, Miller JA, Fist AJ, Gerlach WL, Larkin PJ (2004) RNAi-mediated replacement of morphine with the nonnarcotic alkaloid reticuline in opium poppy. Nat Biotechnol 22(12):1559–1566
• Anthony JR, Anthony LC, Nowroozi F, Kwon G, Newman JD, Keasling JD (2009) Optimization of the mevalonate-based isoprenoid biosynthetic pathway in Escherichia coli for production of the anti-malarial drug precursor amorpha-4, 11-diene. Metab Eng 11:13–19
• Aquil S, Husaini AM, Abdin MZ, Rather GM (2009) Overexpression of the HMG-CoA Reductase Gene Leads to Enhanced Artemisinin Biosynthesis in transgenic Artemisia annua Plants. Planta Med 75:1–6
• Arsenault PR, Wobbe KK, Weathers PJ (2008) Recent advances in artemisinin production through heterologous expression. Curr Med Chem 15:2886
• Arsenault PR, Vail DR, Wobbe KK, Erickson K, Weathers PJ (2010) Reproductive development modulates gene expression and metabolite levels with possible feedback inhibition of artemisinin in A. annua L. Plant Physiol 154:958–968
• Attucci S, Aitken SM, Gulick PJ, Ibrahim RK (1995) Farnesyl pyrophosphate synthase from white lupin: molecular cloning, expression, and purification of the expressed protein. Arch Biochem Biophys 321(2):493–500
• Banyai W, Kirdmanee C, Mii M, Supaibulwatana K (2010) Overexpression of farnesyl pyrophosphate synthase (FPS) gene affected artemisinin content and growth of Artemisia annua L. Plant Cell Tissue Organ. 103:255–265
• Bertea CM, Freije JR, Van der Woude H, Verstappen FW, Perk L, Marquez V, De Kraker JW, Posthumus MA, Jansen BJM, De Groot AE, Franssen MCR, Bouwmeester HJ (2005) Identification of intermediates and enzymes involved in the early steps of artemisinin biosynthesis in Artemisia annua. Planta Med 71:40–47
• Bharel S, Gulati A, Abdin MZ, Srivastava PS, Vishwakarma RA, Jain SK (1998) Enzymatic synthesis of artemisinin from natural and synthetic precursors. J Nat Prod 61:633–636
• Brown GD (2010) The Biosynthesis of Artemisinin (Qinghaosu) and the Phytochemistry of Artemisia annua L. (Qinghao). Molecules 15:7603–7698
• Brown GD, Sy LK (2004) In vivo transformations of dihydroartemisinic acid in Artemisia annua plants. Tetrahedron 60:1139–1159
• Chang MCY, Eachus RA, Trieu W, Ro DK, Keasling JD (2007) Engineering Escherichia coli for production of functionalized terpenoids using plant P450s. Nat Chem Biol 3:274–277
• Chen DH, Liu CJ, Ye HC, Li GF, Liu BY, Meng YL, Chen XY (1999) Ri-mediated transformation of Artemisia annua with a recombinant farnesyl diphosphate synthase gene for artemisinin production. Plant Cell Tissue Organ Cult 57:157–162
• Chen D, Ye H, Li G (2000) Expression of a chimeric farnesyl diphosphate synthase gene in Artemisia annua L. transgenic plants via Agrobacterium tumefaciens-mediated transformation. Plant Sci 155:179–185
• Chen JL, Fang HM, Ji YP, Pu GB, Guo YW, Huang LL, Du ZG, Liu BY, Ye HC, Li GF, Wang H (2011) Artemisinin biosynthesis enhancement in transgenic Artemisia annua plants by downregulation of the b-caryophyllene synthase gene. Planta Med. doi:10.1055/s-0030-1271038
• Covello PS (2008) Making artemisinin. Phytochemistry 69(17): 2881–2885
• De Kraker JW, Scurink M, Franssen MCR, Konig WA, de Groot A, Bouwmeester HJ (2003) Hydroxylation of sesquiterpenes by enzymes from chicory (Cichorium intybus L.) roots. Tetrahedron 59:409–418
• Delabays N, Benakis A, Collet G (1993) Selection and breeding for high artemisinin (Qinghaosu) yielding strains of Artemisia annua. Acta Hort (ISHS). 330:203–208
• Delabays N, Simonnet X, Gaudin M (2001) The genetics of artemisinin content in Artemisia annua L. and the breeding of high yielding cultivars. Curr Med Chem 8:1795–1801
• Delabays N, Darbellay C, Galland N (2002) Variation and heritability of artemisinin content in Artemisia annua L. In: Wright CW (ed) Artemisia. Taylor & Francis, London, pp 197404–209999
• Delourme D, Lacroute F, Karst F (1994) Cloning of an Arabidopsis thaliana cDNA coding for farnesyl diphosphate synthase by functional complementation in yeast. Plant Mol Biol 26(6): 1867–1873
• Duffy PE, Mutabingwa TK (2006) Artemisinin combination therapies. Lancet 367:2037–2039
• Elfawal MA, Towler MJ, Reich NG, Golenbock D, Weathers PJ, Rich SM (2013) Dried whole plant Artemisia annua as an antimalarial therapy. PLoS One 7:e52746. doi:10.1371/journal.pone.0061989
• Farhi M, Marhevka E, Ben-Ari J, Algamas-Dimantov A, Liang Z, Zeevi V, Edelbaum O, Spitzer-Rimon B, Abeliovich H, Schwartz B, Tzfira T, Vainstein A (2011) Generation of the potent anti-malarial drug artemisinin in tobacco. Nat Biotechnol 29(12):1072–1074
• Farmer WR, Liao JC (2001) Precursor balancing for metabolic engineering of lycopene production in Escherichia coli. Biotechnol Prog 17:57–61
• Graham IA, Besser K, Blumer S, Branigan CA, Czechowski T, Elias L, Guterman I, Harvey D, Isaac PG, Khan AM, Larson TR, Li Y, Pawson T, Penfield T, Rae AM, Rathbone DA, Reid S, Ross J et al (2010) The genetic map of Artemisia annua L. identifies loci affecting yield of the antimalarial drug artemisinin. Science 327:328–331
• Han JL, Liu BY, Ye HC, Wang H, Li ZQ, Li GF (2006) Effects of over expression of the endogenous farnesyl diphosphate synthase on the artemisinin content in Artemisia annua L. J Integr Plant Biol 48:482–487
• Harker M, Bramley PM (1999) Expression of prokaryotic 1-deoxy-Dxylulose-5-phosphatases in Escherichia coli increases carotenoid and ubiquinone biosynthesis. FEBS Lett 448:115–119
• Hong GJ, Hu WL, Li JX, Chen XY, Wang LJ (2009) Increased accumulation of artemisinin anthocyanins in Artemisia annua expressing the Arabidopsis blue light receptor CRY1. Plant Mol Biol Rep. doi:10.1007/S11105-008-0088-6
• Jain DC, Mathur AK, Gupta MM, Singh AK, Verma RK, Gupta AP, Kumar S (1996) Isolation of high artemisinin-yielding clones of Artemisia annua. Phytochemistry 6(5):993–1001
• Jing FY, Zhang L, Li MY, Tang KX (2008) Over-expressing cyp71avl and cpr genes enhances artemisinin content in Artemisa annua L. J Agr Sci Tech Iran 10(3):64–70
• Kajiwara S, Fraser PD, Kondo K, Misawa N (1997) Expression of an exogenous isopentenyl diphosphate isomerase gene enhances isoprenoid biosynthesis in Escherichia coli. Biochem J 324:421–426
• Korth KL, Jaggard DAW, Dixon RA (2000) Development and light regulated post translational control of 3-hydroxy 3-methylglutary coenzyme A reductase levels in potato. Plant J 23:507–516
• Kudakasseril GJ, Lam L, Staba EJ (1987) Effect of sterol inhibitors on the incorporation of 14C-isopentylpyrophosphate into artemisinin by a cell-free system from Artemisia annua tissue cultures and plants. Planta Med 53:280–284
• Lenihan JR, Tsuruta H, Diola D, Renninger NS, Regentin R (2008) Developing an industrial artemisinic acid fermentation process to support the cost-effective production of antimalarial artemisininbased combination therapies. Biotechnol Prog 24:1026–1032
• Li Y, Huang H, Wu YL (2006) Ginghaosu (artemisinin)—a fantastic antimalarial drug from a traditional Chinese medicine. In: Liang X-T, Fang WS (Eds.) Medicinal chemistry of bioactive natural products. Wiley Interscience, Hoboken, pp 183–256
• Liu J, Ni M, Fan J, Tu Y, Wu Z, Wu Y, Zhou W (1979) Structure and reaction of Qinghaosu (in Chinese). Acta Chim Sinica 37:129–143
• Liu CZ, Guo C, Wang YC, Ouyang F (2002) Effect of light irradiation on hairy root growth and artemisinin biosynthesis of Artemisia annua L. Process Biochem 38:581–585
• Liu Y, Ye HC, Wang H, Li GF (2003) Molecular cloning, Escherichia coli expression and genomic organization of squalene synthase gene from Artemisia annua. Acta Bot Sin 45:608–613
• Liu C, Zhao Y, Wang Y (2006) Artemisinin: current state and perspectives for biotechnological production of an antimalarial drug. Appl Mcrobiol Biotechnol 72:11–20
• Liu B, Wang H, Du Z, Li G, Ye H (2011) Metabolic engineering of artemisinin biosynthesis in Artemisia annua L. Plant Cell Rep 30(5):689–694
• Ma C, Wang H, Lu X, Xu G, Liu B (2008) Metabolic fingerprinting investigation of Artemisia annua L. in different stages of development by gas chromatography and gas chromatography– mass spectrometry. J Chromatogr A 1186:412–419
• Ma C, Wang HH, Lu X, Wang H, Xu G, Liu B (2009a) Terpenoid metabolic profiling analysis of transgenic Artemisia annua L. by comprehensive two-dimensional gas chromatography time-offlight mass spectrometry. Metabolomics 5:497–506
• Ma D, Pu G, Lei C, Ma L, Wang H, Guo Y, Chen J, Du Z, Li G, Ye H, Liu B (2009b) Isolation and characterization of AaWRKY1, an Artemisia annua transcription factor that regulates the amorpha- 4,11-diene synthase gene, a key gene of artemisinin biosynthesis. Plant Cell Physiol 50:2146–2161
• Maes L, Van Nieuwerburgh FCW, Zhang YS, Reed DW, Pollier J, Casteele S, Inze D, Covello PS, Deforce DLD, Goossens A (2011) Dissection of the phytohormonal regulation of trichome formation and biosynthesis of the antimalarial compound artemisinin in Artemisia annua plants. New Phytol 189:176–189
• Martin VJJ, Pitera DJ, Withers ST, Newman JD, Keasling JD (2003) Engineering a mevalonate pathway in Escherichia coli for production of terpenoids. Nature Biotech 7:796–802
• Matsushita Y, Kang WY, Charlwood BV (1996) Cloning and analysis of a cDNA encoding farnesyl diphosphate synthase from Artemisia annua. Gene 172:207–209
• Mercke P, Bengtsson M, Bouwmeester HJ, Posthumus MA, Brodelius PE (2000) Molecular cloning, expression, and characterization of amorpha-4,11-diene synthase, a key enzyme of artemisinin biosynthesis in Artemisia annua L. Arch Biochem Biophys 381:173–180
• Nafis T, Akmal M, Mauji Ram, Alam P, Ahlawat S, Mohammad A, Abdin MZ (2011) Enhancement of artemisinin content by constitutive expression of HMG CoA Reductase gene in high yielding strain of Artemisia annua L. Plant Biotechnol Rep. 5:53–60
• Nair MSR, Basile DV (1993) Bioconversion of arteannuin B to artemisinin. J Nat Prod 56:1559–1566
• Nair P, Misra A, Singh A, Shukla AK, Gupta MM et al (2013) Differentially expressed genes during contrasting growth stages of Artemisia annua for artemisinin content. PLoS ONE 8(4):e60375. doi:10.1371/journal.pone.0060375
• Nguyen KT, Arsenault PR, Weathers PJ (2011) Trichomes +roots + ROS = artemisinin: regulating artemisinin biosynthesis in Artemisia annua L. In Vitro Cell Dev Biol Plant 47:329–338
• Ogita S, Uefuji H, Morimoto M, Sano H (2004) Application of RNAi to confirm theobromine as the major intermediate for caffeine biosynthesis in coffee plants with potential for construction of decaffeinated varieties. Plant Mol Biol 54(6):931–941
• Olsson ME, Olofsson LM, Lindahl AL, Lundgren A, Brodelius M, Brodelius PE (2009) Localization of enzymes of artemisinin biosynthesis to the apical cells of glandular secretory trichomes of Artemisia annua L. Phytochemistry 70:1123–1128
• Paddon CJ, Westfall PJ, Pitera DJ, Benjamin K, Fisher K, McPhee D, Leavell MD, Tai A, Main A, Eng D, Polichuk DR et al (2013) High-level semisynthetic production of the potent antimalarial artemisinin. Nature 496:528–532
• Paul S, Khanuja SP, Shasany AK, Gupta MM, Darokar MP, Saikia D, Gupta AK (2010) Enhancement of artemisinin content through four cycles of recurrent selection with relation to heritability, correlation and molecular marker in Artemisia annua L. Planta Med 76:1468–1472
• MaujiRam, Khan MA, Jha P, Khan S, Kiran U, Ahmad MM, Javed S, Abdin MZ (2010) HMG-CoA reductase limits artemisinin biosynthesis and accumulation in Artemisia annua L. plants. Acta Physiol Plant 32:859–866
• Ro DK, Paradise EM, Ouellet M, Fisher KJ, Newman KL, Ndungu JM, Ho KA, Eachus RA, Ham TS, Kirby J, Chang MC, Withers ST, Shiba Y, Sarpong R, Keasling JD (2006) Production of the antimalarial drug precursor artemisinic acid in engineered yeast. Nature 440:940–943
• Rydén AM, Ruyter-Spira C, Quax WJ, Osada H, Muranaka T, Kayser O, Bouwmeester H (2010) The molecular cloning of dihydroartemisinic aldehyde reductase and its implication in artemisinin biosynthesis in Artemisia annua. Planta Med 76(15):1778–1783
• Sa G, Mi M, He-Chun Y, Ben-Ye L, Guo-feng L, Kang C (2001) Effects of ipt gene expression on the physiological and chemical characteristics of Artemisia annua L. Plant Sci 160:691–698
• Schramek N, Wang H, Romisch-Margl W, Keil B, Radykewicz T, Winzenhorlein B, Beerhues L, Bacher A, Rohdich F, Gershenzon J, Liu B, Eisenreich W (2010) Artemisinin biosynthesis in growing plants of Artemisia annua. A 13CO2 study. Phytochemistry 71:179–187
• Shen Q, Chen YF, Wang T, Wu SY, Lu X, Zhang L, Zhang FY, Jiang WM, Wang GF, Tang KX (2012) Overexpression of the cytochrome P450 monooxygenase (cyp71av1) and cytochrome P450 reductase (cpr) genes increased artemisinin content in Artemisia annua (Asteraceae). Genet Mol Res 11:3298–3309
• Tang KX, Shen Q, Chen YF, Wang T, Wu SY, Lu X (2012b) Overexpression DBR2 gene increased artemisinin content in Artemisia annua L. Shanghai Jiao Tong University, China. Patent CN201210014227.2
• Tang KX, Chen YF, Shen Q, Wang T, Wu SY, Wang GF (2012c) Overexpression ALDH1 gene increased artemisinin content in Artemisia annua L. Shanghai Jiao Tong University, China. Patent CN201210014242.7
• Teoh KH, Polichuk DR, Reed DW, Nowak G, Covello PS (2006) 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 580:1411–1416
• Teoh KH, Polichuk DR, Reed DW, Nowak G, Covello PS (2009) Molecular cloning of an aldehyde dehydrogenase implicated in artemisinin biosynthesis in Artemisia annua. Botany. 87: 635–642
• Towler MJ, Weathers PJ (2007) Evidence of artemisinin production from IPP stemming from both the mevalonate and the nonmevalonate pathways. Plant Cell Rep 26(12):2129–2136
• Townsend T, Segura V, Chigeza G, Penfield T, Rae A, Harvey D, Bowles D, Graham IA (2013) The use of combining ability analysis to identify elite parents for Artemisia annua F1 hybrid production. PLoS One 8(4):e61989. doi:10.1371/journal.pone. 0061989
• Tsuruta H, Paddon CJ, Eng D, Lenihan JR, Horning T, Anthony LC, Regentin R, Keasling JD, Renninger NS, Newman JD (2009) High-level production of amorpha-4,11-diene, a precursor of the antimalarial agent artemisinin, in Escherichia coli. PLoS One 4:e4489
• Wallaart TE, Bouwmeester HJ, Hille J, Poppinga L, Maijers NC (2001) Amorpha-4, 11-diene synthase: cloning and functional expression of a key enzyme in the biosynthetic pathway of the novel antimalarial drug artemisinin. Planta 212:460–465
• Wang YY, Jing FY, Yu SY, Chen YF, Wang T, Liu P, Wang GF, Sun XF, Tang KX (2011a) Co-overexpression of the HMGR and FPS genes enhances artemisinin content in Artemisia annua L. J Med Plants Res 5:3396–3403
• Wang YY, Yang K, Deng T, Huang R, Wang B, Wang GF, Sun XF, Tang KX (2011b) Cloning and characterization of trichome specific promoter of cpr71av1 gene involved in artemisinin biosynthesis in Artemisia annua L. Mol Biol 145:751–758
• Wang HZ, Han JL, Kanagarajan S, Lundgren A, Brodelius PE (2013) Trichome-specific expression of the amorpha-4,11-diene 12-hydroxylase (cyp71av1) gene, encoding a key enzyme of artemisinin biosynthesis in Artemisia annua, as reported by a promoter-GUS fusion. Plant Mol Biol 81:119–138
• Weathers P, Elkholy S, Wobbe KK (2006) Invited review: artemisinin: the biosynthetic pathway and its regulation in Artemisia annua, a terpenoid-rich species. In Vitro Cell Dev Biol Plant 42:309–317
• Weathers PJ, Jordan NJ, Lasin P, Towler MJ (2014) Simulated digestion of dried leaves of Artemisia annua consumed as a treatment (pACT) for malaria. J Ethnopharmacol 151(2):858–863
• Westfall PJ, Pitera DJ, Lenihan JR, Eng D, Woolard FX, Regentin R, Horning T, Tsuruta H, Melis DJ, Owens A, Fickes S, Diola D, Benjamin KR, Keasling JD, Leavell MD, McPhee DJ, Renninger NS, Newman JD, Paddon CJ (2012) Production of amorphadiene in yeast, and its conversion to dihydroartemisinic acid, precursor to the antimalarial agent artemisinin. Proc Natl Acad Sci 109:E111–E118
• White NJ (2008) Qinghaosu (artemisinin): the price of success. Science 320:330–334
• Woerdenbag HJ, Jos FJ, van Uden W, Pras N, Malingre TM, Alfermann AW (1993) Production of the new antimalarial drug artemisinin in shoot cultures of Artemisia annua L. Plant Cell, Tissue Organ Cult 32:247–257
• Xiang LE, Zeng LX, Yuan Y, Chen M, Wang F, Liu XQ, Zeng LJ, Lan XZ, Liao ZH (2012) Enhancement of artemisinin biosynthesis by overexpressing dxr, cyp71av1 and cpr in the plants of Artemisia annua L. Plant Omics J 5:503–507
• Yu ZX, Li JX, Yang CQ, Hu WL, Wang LJ, Chen XY (2012) The jasmonate-responsive AP2/ERF transcription factors AaERF1 and AaERF2 positively regulate artemisinin biosynthesis in Artemisia annua L. Mol Plant 5(2):353–365
• Zhang YS, Teoh KH, Reed DW, Maes L, Goossens A, Olson DJ, Ross ARS, Covello PS (2008) The Molecular Cloning of Artemisinic Aldehyde 1–1(13) reductase and Its Role in Glandular Trichomedependent Biosynthesis of Artemisinin in Artemisia annua. J Bio Chem 283:21501–21508
• Zhang L, Jing F, Li F, Li M, Wang Y, Wang G, Sun X, Tang K (2009) 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 52:199–207
• Zhang F, Lu X, Lv Z, Zhang L, Zhu M, Jiang W, Wang G, Sun X, Tang K (2013) Overexpression of the Artemisia orthologue of ABA receptor, AaPYL9, enhances ABA sensitivity and improves artemisinin content in Artemisia annua L. PLoS One 8(2):e56697
• Zhu C, Cook SP (2012) A concise synthesis of (?)—artemisinin. J Am Chem Soc 134:13577–13579

Identyfikatory

DOI

10.1007/s11738-015-177-5