production of artemisinic acid using engineered yeast journal club i 7 th july 09 david roche...
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Production of Artemisinic Production of Artemisinic acid using engineered acid using engineered
yeastyeast
Journal Club I7th July 09
David RocheCharles Fracchia
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SummarySummary Introduction
Results
Concept of feedback
Discussion
How is it relevant to SB?
Conclusions
Materials and MethodsIdentifying the genes involved in Artemisinin production
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IntroductionIntroductionArtemisinin is anti-malarial compound
Currently extracted from the wormwood plant – but not efficient or cheap enough
Copied the biosynthetic pathways into the yeast
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Materials and MethodsMaterials and Methods
Green: engineered pathways
Blue: directly upregulated
Purple: indirectly upregulated
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Materials and MethodsMaterials and Methods
Increased FPP production by upregulating FPP synthases and downregulating to convertases
Introduced ADSCloned P450
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M&M: Identifying the ADS M&M: Identifying the ADS genesgenes
They supposed that the enzymes shown in green shared common ancestor enzymes
Compared the genes using BLAST and identified one P450 gene with high homology
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ResultsResults
5x
2x
50%
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The concept of feedback The concept of feedback inhibition/activationinhibition/activation
Metabolic flux relies on regulation
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DiscussionDiscussion Increase in yield and decrease in
production costs
General principle can be applied to production of other compounds, e.g. Taxol – an anti cancer drug, which is normally extracted from the Pacific yew tree.
Good example of metabolic engineering to give a useful product.
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DiscussionDiscussionLaborious process of specially
engineering each step.
Not necessarily easily reproducible. To re-engineer for other compounds, must go ‘back to the drawing board.’
Yield optimization and industrial scale-up still required to reduce prices significantly below their current level.
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How is it relevant to SB?How is it relevant to SB?Previous strategies in metabolic engineering
seem more of an art with experimentation by trial-and-error.
Keasling approach to the problem was more in line with the principles of Synthetic Biology, using a logical approach for the design.
Used computational modelling to investigate the most efficient mRNA sequence for maximal compound production
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ConclusionsConclusions Materials and Methods
Results
Concept of feedback
Discussion
Duplicate genes
Knockout genes
Genetic insertion
50% increase for duplication
2x increase for knockout
5x increase for gene insertion
Products of a reaction can control
their own conversion
Engineered approach to metabolic engineering.
Basic method can be applied to production of other compounds.