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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 transcript levels of ADS, CPR, CYP71AV1, FPS, and HMGR genes in suspension-cultured cells of A. annua. (1): control group; (2): DHAA (25 mg/L) treated group, UBC was used as the internal standard. The band intensities were quantified by Image J software, and the relative expression was the ratio of intensity of control group to that of DHAA group.
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fig5: Effect of DHAA on the transcript levels of ADS, CPR, CYP71AV1, FPS, and HMGR genes in suspension-cultured cells of A. annua. (1): control group; (2): DHAA (25 mg/L) treated group, UBC was used as the internal standard. The band intensities were quantified by Image J software, and the relative expression was the ratio of intensity of control group to that of DHAA group.

Mentions: As shown in Figure 5, ADS, CYP71AV1, and FPS transcript levels in cultured cells of A. annua were upregulated by the addition of DHAA, in which the transcript abundances were 3.19-, 7.21-, and 2.04-fold higher than those of the control group, respectively. While CPR and HMGR transcripts were detected both in the control group and in the DHAA-treated group, the transcript abundances for these two groups appeared little different.


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 transcript levels of ADS, CPR, CYP71AV1, FPS, and HMGR genes in suspension-cultured cells of A. annua. (1): control group; (2): DHAA (25 mg/L) treated group, UBC was used as the internal standard. The band intensities were quantified by Image J software, and the relative expression was the ratio of intensity of control group to that of DHAA group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig5: Effect of DHAA on the transcript levels of ADS, CPR, CYP71AV1, FPS, and HMGR genes in suspension-cultured cells of A. annua. (1): control group; (2): DHAA (25 mg/L) treated group, UBC was used as the internal standard. The band intensities were quantified by Image J software, and the relative expression was the ratio of intensity of control group to that of DHAA group.
Mentions: As shown in Figure 5, ADS, CYP71AV1, and FPS transcript levels in cultured cells of A. annua were upregulated by the addition of DHAA, in which the transcript abundances were 3.19-, 7.21-, and 2.04-fold higher than those of the control group, respectively. While CPR and HMGR transcripts were detected both in the control group and in the DHAA-treated group, the transcript abundances for these two groups appeared little different.

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