Production of Artemisinic acid using engineered yeast Journal Club I 7 th July 09 David Roche...
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Transcript of Production of Artemisinic acid using engineered yeast Journal Club I 7 th July 09 David Roche...
Production of Artemisinic Production of Artemisinic acid using engineered acid using engineered
yeastyeast
Journal Club I7th July 09
David RocheCharles Fracchia
SummarySummary Introduction
Results
Concept of feedback
Discussion
How is it relevant to SB?
Conclusions
Materials and MethodsIdentifying the genes involved in Artemisinin production
IntroductionIntroductionArtemisinin is anti-malarial compound
Currently extracted from the wormwood plant – but not efficient or cheap enough
Copied the biosynthetic pathways into the yeast
Materials and MethodsMaterials and Methods
Green: engineered pathways
Blue: directly upregulated
Purple: indirectly upregulated
Materials and MethodsMaterials and Methods
Increased FPP production by upregulating FPP synthases and downregulating to convertases
Introduced ADSCloned P450
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
ResultsResults
5x
2x
50%
The concept of feedback The concept of feedback inhibition/activationinhibition/activation
Metabolic flux relies on regulation
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.
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.
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
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.