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Inducing effect of dihydroartemisinic acid in the biosynthesis of artemisinins with cultured cells of Artemisia annua by enhancing the expression of genes.

Zhu J, Yang J, Zeng Z, Zhang W, Song L, Wen W, Yu R - ScientificWorldJournal (2014)

Bottom Line: The structures of four DHAA metabolites, namely, dihydro-epi-deoxyarteannuin B, arteannuin I, arteannuin K, and 3-β-hydroxy-dihydro-epi-deoxyarteannuin B, were elucidated by physicochemical and spectroscopic analyses.The correlation between gene expression and ART content was investigated.The results of RT-PCR showed that DHAA could up-regulate expression of amorpha-4,11-diene synthase gene (ADS), amorpha-4,11-diene C-12 oxidase gene (CYP71AV1), and farnesyl diphosphate synthase gene (FPS) (3.19-, 7.21-, and 2.04-fold higher than those of control group, resp.), which indicated that biosynthesis processes from DHAA to ART were enzyme-mediated.

View Article: PubMed Central - PubMed

Affiliation: Biotechnological Institute of Chinese Materia Medica, Jinan University, Guangzhou 510632, China.

ABSTRACT
Artemisinin has been used in the production of "artemisinin combination therapies" for the treatment of malaria. Feeding of precursors has been proven to be one of the most effective methods to enhance artemisinin production in plant cultured cells. At the current paper, the biosynthesis of artemisinin (ART) and its four analogs from dihydroartemisinic acid (DHAA) in suspension-cultured cells of Artemisia annua were investigated. ARTs were detected by HPLC/GC-MS and isolated by various chromatography methods. The structures of four DHAA metabolites, namely, dihydro-epi-deoxyarteannuin B, arteannuin I, arteannuin K, and 3-β-hydroxy-dihydro-epi-deoxyarteannuin B, were elucidated by physicochemical and spectroscopic analyses. The correlation between gene expression and ART content was investigated. The results of RT-PCR showed that DHAA could up-regulate expression of amorpha-4,11-diene synthase gene (ADS), amorpha-4,11-diene C-12 oxidase gene (CYP71AV1), and farnesyl diphosphate synthase gene (FPS) (3.19-, 7.21-, and 2.04-fold higher than those of control group, resp.), which indicated that biosynthesis processes from DHAA to ART were enzyme-mediated.

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Related in: MedlinePlus

Biosynthesis pathway of ART.
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fig1: Biosynthesis pathway of ART.

Mentions: It is well known that biosynthesis of ART includes three stages, that is, the formation of farnesyl pyrophosphate (FPP) from acetyl-CoA, the synthesis of sesquiterpene, and the formation of ART via lactonization (Figure 1). Farnesyl pyrophosphate synthase (FPS) catalyzes condensation of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) to produce FPP, which is the starting point of a large variety of important isoprenoid end products, including ART [9]. The cyclization of FPP to generate amorpha-4,11-diene catalyzed by amorpha-4,11-diene synthase (ADS) has been suggested as the first committed and limiting step in ART biosynthesis [10]. CYP71AV1, a cytochrome P450, could oxidize amorpha-4,11-diene to generate artemisinic aldehyde intermediates, which might be further catalyzed to dihydroartemisinic acid (DHAA) by artemisinic aldehyde Δ11(13) reductase (Dbr2) and aldehyde dehydrogenase (Aldh1) [11–13]. DHAA was supposed to be converted chemically into ART by an oxygen-mediated photochemical oxidation in vitro, which was a nonenzymatic process [14, 15]. Some experimental results also demonstrated that enzymatic catalysis or both enzymatic catalysis and autooxidation might occurr in the pathway from DHAA to ART in A. annua [16].


Inducing effect of dihydroartemisinic acid in the biosynthesis of artemisinins with cultured cells of Artemisia annua by enhancing the expression of genes.

Zhu J, Yang J, Zeng Z, Zhang W, Song L, Wen W, Yu R - ScientificWorldJournal (2014)

Biosynthesis pathway of ART.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4127257&req=5

fig1: Biosynthesis pathway of ART.
Mentions: It is well known that biosynthesis of ART includes three stages, that is, the formation of farnesyl pyrophosphate (FPP) from acetyl-CoA, the synthesis of sesquiterpene, and the formation of ART via lactonization (Figure 1). Farnesyl pyrophosphate synthase (FPS) catalyzes condensation of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) to produce FPP, which is the starting point of a large variety of important isoprenoid end products, including ART [9]. The cyclization of FPP to generate amorpha-4,11-diene catalyzed by amorpha-4,11-diene synthase (ADS) has been suggested as the first committed and limiting step in ART biosynthesis [10]. CYP71AV1, a cytochrome P450, could oxidize amorpha-4,11-diene to generate artemisinic aldehyde intermediates, which might be further catalyzed to dihydroartemisinic acid (DHAA) by artemisinic aldehyde Δ11(13) reductase (Dbr2) and aldehyde dehydrogenase (Aldh1) [11–13]. DHAA was supposed to be converted chemically into ART by an oxygen-mediated photochemical oxidation in vitro, which was a nonenzymatic process [14, 15]. Some experimental results also demonstrated that enzymatic catalysis or both enzymatic catalysis and autooxidation might occurr in the pathway from DHAA to ART in A. annua [16].

Bottom Line: The structures of four DHAA metabolites, namely, dihydro-epi-deoxyarteannuin B, arteannuin I, arteannuin K, and 3-β-hydroxy-dihydro-epi-deoxyarteannuin B, were elucidated by physicochemical and spectroscopic analyses.The correlation between gene expression and ART content was investigated.The results of RT-PCR showed that DHAA could up-regulate expression of amorpha-4,11-diene synthase gene (ADS), amorpha-4,11-diene C-12 oxidase gene (CYP71AV1), and farnesyl diphosphate synthase gene (FPS) (3.19-, 7.21-, and 2.04-fold higher than those of control group, resp.), which indicated that biosynthesis processes from DHAA to ART were enzyme-mediated.

View Article: PubMed Central - PubMed

Affiliation: Biotechnological Institute of Chinese Materia Medica, Jinan University, Guangzhou 510632, China.

ABSTRACT
Artemisinin has been used in the production of "artemisinin combination therapies" for the treatment of malaria. Feeding of precursors has been proven to be one of the most effective methods to enhance artemisinin production in plant cultured cells. At the current paper, the biosynthesis of artemisinin (ART) and its four analogs from dihydroartemisinic acid (DHAA) in suspension-cultured cells of Artemisia annua were investigated. ARTs were detected by HPLC/GC-MS and isolated by various chromatography methods. The structures of four DHAA metabolites, namely, dihydro-epi-deoxyarteannuin B, arteannuin I, arteannuin K, and 3-β-hydroxy-dihydro-epi-deoxyarteannuin B, were elucidated by physicochemical and spectroscopic analyses. The correlation between gene expression and ART content was investigated. The results of RT-PCR showed that DHAA could up-regulate expression of amorpha-4,11-diene synthase gene (ADS), amorpha-4,11-diene C-12 oxidase gene (CYP71AV1), and farnesyl diphosphate synthase gene (FPS) (3.19-, 7.21-, and 2.04-fold higher than those of control group, resp.), which indicated that biosynthesis processes from DHAA to ART were enzyme-mediated.

Show MeSH
Related in: MedlinePlus