Combination of ERG9 Repression and Enzyme Fusion Technology for Improved Production of Amorphadiene in Saccharomyces cerevisiae
The yeast strain (Saccharomyces cerevisiae) MTCC 3157 was selected for combinatorial biosynthesis of plant sesquiterpene amorpha-4,11-diene. Our main objective was to overproduce amorpha 4-11-diene, which is a key precursor molecule of artemisin...
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| Language: | English |
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Wiley
2013-01-01
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| Series: | Journal of Analytical Methods in Chemistry |
| Online Access: | http://dx.doi.org/10.1155/2013/140469 |
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| author | Rama Raju Baadhe Naveen Kumar Mekala Sreenivasa Rao Parcha Yalavarthy Prameela Devi |
| author_facet | Rama Raju Baadhe Naveen Kumar Mekala Sreenivasa Rao Parcha Yalavarthy Prameela Devi |
| author_sort | Rama Raju Baadhe |
| collection | DOAJ |
| description | The yeast strain (Saccharomyces cerevisiae) MTCC 3157
was selected for combinatorial biosynthesis of plant sesquiterpene amorpha-4,11-diene.
Our main objective was to overproduce amorpha 4-11-diene, which is a key precursor molecule of
artemisinin (antimalarial drug) produced naturally in plant Artemisia annua through
mevalonate pathway. Farnesyl diphosphate (FPP) is a common intermediate metabolite of a variety
of compounds in the mevalonate pathway of yeast and leads to the production of ergosterols,
dolichol and ubiquinone, and so forth. In our studies, FPP converted to amorphadiene (AD) by
expressing heterologous amorphadiene synthase (ADS) in yeast. First,
ERG9 (squalane synthase) promoter of yeast was replaced with repressible
methionine (MET3) promoter by using bipartite gene fusion method. Further to overcome the loss of the
intermediate FPP through competitive pathways in yeast, fusion protein technology was adopted
and farnesyldiphosphate synthase (FPPS) of yeast has been coupled with amorphadiene
synthase (ADS) of plant origin (Artemisia annua L.) where amorphadiene
production was improved by 2-fold (11.2 mg/L) and 4-fold (25.02 mg/L) in yeast strains
YCF-002 and YCF-005 compared with control strain YCF-AD (5.5 mg/L), respectively. |
| format | Article |
| id | doaj-art-1efa2728bc944b5a8bb355565dd9f033 |
| institution | Kabale University |
| issn | 2090-8865 2090-8873 |
| language | English |
| publishDate | 2013-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Analytical Methods in Chemistry |
| spelling | doaj-art-1efa2728bc944b5a8bb355565dd9f0332025-02-03T01:30:15ZengWileyJournal of Analytical Methods in Chemistry2090-88652090-88732013-01-01201310.1155/2013/140469140469Combination of ERG9 Repression and Enzyme Fusion Technology for Improved Production of Amorphadiene in Saccharomyces cerevisiaeRama Raju Baadhe0Naveen Kumar Mekala1Sreenivasa Rao Parcha2Yalavarthy Prameela Devi3Department of Biotechnology, National Institute of Technology, Warangal 506004, IndiaDepartment of Biotechnology, National Institute of Technology, Warangal 506004, IndiaDepartment of Biotechnology, National Institute of Technology, Warangal 506004, IndiaDepartment of Zoology, Kakatiya University, Warangal, Andhra Pradesh 506009, IndiaThe yeast strain (Saccharomyces cerevisiae) MTCC 3157 was selected for combinatorial biosynthesis of plant sesquiterpene amorpha-4,11-diene. Our main objective was to overproduce amorpha 4-11-diene, which is a key precursor molecule of artemisinin (antimalarial drug) produced naturally in plant Artemisia annua through mevalonate pathway. Farnesyl diphosphate (FPP) is a common intermediate metabolite of a variety of compounds in the mevalonate pathway of yeast and leads to the production of ergosterols, dolichol and ubiquinone, and so forth. In our studies, FPP converted to amorphadiene (AD) by expressing heterologous amorphadiene synthase (ADS) in yeast. First, ERG9 (squalane synthase) promoter of yeast was replaced with repressible methionine (MET3) promoter by using bipartite gene fusion method. Further to overcome the loss of the intermediate FPP through competitive pathways in yeast, fusion protein technology was adopted and farnesyldiphosphate synthase (FPPS) of yeast has been coupled with amorphadiene synthase (ADS) of plant origin (Artemisia annua L.) where amorphadiene production was improved by 2-fold (11.2 mg/L) and 4-fold (25.02 mg/L) in yeast strains YCF-002 and YCF-005 compared with control strain YCF-AD (5.5 mg/L), respectively.http://dx.doi.org/10.1155/2013/140469 |
| spellingShingle | Rama Raju Baadhe Naveen Kumar Mekala Sreenivasa Rao Parcha Yalavarthy Prameela Devi Combination of ERG9 Repression and Enzyme Fusion Technology for Improved Production of Amorphadiene in Saccharomyces cerevisiae Journal of Analytical Methods in Chemistry |
| title | Combination of ERG9 Repression and Enzyme Fusion Technology for Improved Production of Amorphadiene in Saccharomyces cerevisiae |
| title_full | Combination of ERG9 Repression and Enzyme Fusion Technology for Improved Production of Amorphadiene in Saccharomyces cerevisiae |
| title_fullStr | Combination of ERG9 Repression and Enzyme Fusion Technology for Improved Production of Amorphadiene in Saccharomyces cerevisiae |
| title_full_unstemmed | Combination of ERG9 Repression and Enzyme Fusion Technology for Improved Production of Amorphadiene in Saccharomyces cerevisiae |
| title_short | Combination of ERG9 Repression and Enzyme Fusion Technology for Improved Production of Amorphadiene in Saccharomyces cerevisiae |
| title_sort | combination of erg9 repression and enzyme fusion technology for improved production of amorphadiene in saccharomyces cerevisiae |
| url | http://dx.doi.org/10.1155/2013/140469 |
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