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Biosynthesis of Taxol in E. coli
Duke iGEM 2016
What is Taxol? Why does it matter?
Bark of Pacific Yew Tree Chemotherapy Medication
Project OverviewHow can we use synthetic biology to make
taxol treatment more accessible?
Taxol Production Issues
Taking from nature and chemical synthesis is Extremely inefficient, costly, etc.
Currently using plants and fermentation of plant cells
Our Goal: Biosynthesis of Taxol in E. coli
If we can produce it via bacteria, Time savingCost saving Life saving
Methods and Experimentation
Phosphate Promoter
Xba1
Benzoate-CoA Ligase GeneRBShomology homology
Spe1
homology
Design of G-Block Inserts
Phosphate Promoter
Xba1
Benzoate-CoA Ligase GeneRBShomology homology
Spe1
homology
Design of G-Block Inserts
Kinetic AssayShades of red indicate varying
concentrations of benzoic acidMagenta is the empty vector
control with a benzoic acid concentration of 0.5 mM
Orange is lack of ATPBlue a lack of CoA, and Green a lack of benzoic acidPurple box represents badA
enzyme Green box represents EV
Results
Discussion
Activity Number
Describes the amount of substrates used per binding site per minute
Characterizes the enzyme based on how active it is
Allows for identification of bottleneck enzymes in a pathway
Controls for side reaction and endogenous reactions
Comparisons of different concentrations of substrates
Future Plans
The rest of the enzymes need to be characterized Each must be optimized to avoid bottlenecks in the
process A Golden Gate Assembly should be constructed so
that taxol can be produced via fermentation of bacteria cells
Outside the Lab!
Optimization of biosynthesis would drastically decrease the price of Taxol
Examined social policy to see how such a decrease would help
Results of our analysis Scientists and policymakers will have to
address more systemic issues in the current system to force prices down
Just the science isn’t enough
Effect on Health Care?
Patent on taxol?
Brainstorming marketability of project
Potential patentability of genetically-modified bacteria
Important for incentivizing pharmaceutical companies
They would want exclusive right to the invention
Community Outreach
Helping Gaston Day High School
Henna Saqibo, Lilith Tang, Parth Patel, Wiliam Henry
● Advised team on how to proceed with Arabinose Induction
● Sent them specialized E.coli strain● Helped them troubleshoot● Went and talked to them about their plans, and
how to move forward ● Analyzed a colicin sample transformed in the
pSB1C3 backbone
Synthetic Biology Lecture at Duke
“Should we engineer the Mosquito?” forum
Michael Lynch, M.D., Ph.D
Duke Biomedical Engineering
Building with Bio
Museum of Life and ScienceRaleigh, North Carolina
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