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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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Effect of DHAA on the yield of ART.
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fig4: Effect of DHAA on the yield of ART.

Mentions: To determine the optimal biosynthesis conditions, different concentrations of DHAA and the coculture time were investigated. The results indicated that there was no ART detected in the two control groups and DHAA (50 mg/L) treated group (Figure 4). High amount of DHAA (50 mg/L) showed great toxicity to the culture cells, which made the cells dead from the first day. The highest yield (237.5 µg/L) occurred on day 2 after the coculture in DHAA (25 mg/L) treated group. Therefore, the optimal condition for ART production was shown as follow: precultured time for cells of A. annua: 13 days; concentration of DHAA: 25 mg/L; cocultured time: 2 days.


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)

Effect of DHAA on the yield of ART.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig4: Effect of DHAA on the yield of ART.
Mentions: To determine the optimal biosynthesis conditions, different concentrations of DHAA and the coculture time were investigated. The results indicated that there was no ART detected in the two control groups and DHAA (50 mg/L) treated group (Figure 4). High amount of DHAA (50 mg/L) showed great toxicity to the culture cells, which made the cells dead from the first day. The highest yield (237.5 µg/L) occurred on day 2 after the coculture in DHAA (25 mg/L) treated group. Therefore, the optimal condition for ART production was shown as follow: precultured time for cells of A. annua: 13 days; concentration of DHAA: 25 mg/L; cocultured time: 2 days.

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