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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

GC-MS detection of DHAA metabolites in suspension-cultured cells of A. annua. (I): suspension-cultured cells with DHAA; (II): suspension-cultured cells without DHAA; (III): media with DHAA but without suspension-cultured cells.
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Related In: Results  -  Collection


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fig2: GC-MS detection of DHAA metabolites in suspension-cultured cells of A. annua. (I): suspension-cultured cells with DHAA; (II): suspension-cultured cells without DHAA; (III): media with DHAA but without suspension-cultured cells.

Mentions: Biosynthesis products of DHAA were found in EtOAc extraction of group I (Figure 2). Compared the GC-MS spectra of group I and group II, at least 19 new peaks (a~s) appeared, which showed that enzyme-catalyzed reaction or autooxidation likely occurred in group I.


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)

GC-MS detection of DHAA metabolites in suspension-cultured cells of A. annua. (I): suspension-cultured cells with DHAA; (II): suspension-cultured cells without DHAA; (III): media with DHAA but without suspension-cultured cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig2: GC-MS detection of DHAA metabolites in suspension-cultured cells of A. annua. (I): suspension-cultured cells with DHAA; (II): suspension-cultured cells without DHAA; (III): media with DHAA but without suspension-cultured cells.
Mentions: Biosynthesis products of DHAA were found in EtOAc extraction of group I (Figure 2). Compared the GC-MS spectra of group I and group II, at least 19 new peaks (a~s) appeared, which showed that enzyme-catalyzed reaction or autooxidation likely occurred in group I.

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