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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