1. Background
2. Building the horse - Engineering a novel strain for antibiotics-free selection
3. The army inside - Toluene-4-Monooxygenase (T4MO)
Toluene-4-Monooxygenase (T4MO) is a three-component enzyme system that is naturally able to degrade a variety of hydrocarbons. In particular, its ability to oxidize and reduce small aromatic compounds has caught the attention of scientists looking for biological means of producing certain useful organic chemicals for industrial use, resulting in extensive research on modifying its activity and affinity.
For our project, a mutated form of T4MO (double mutations at G103L & A107G) was chosen as the candidate for indole degradation. While there were several possible candidate enzymes, this particular one was known to convert almost 50% of the indole into 7-hydroxyindole (7-HI). This is of great interest to us as 7-HI has been documented to inhibit biofilm formation in Enterohemorrhagic E. coli, and quite possibly for other strains of E. coli as well. Biofilm formation is a major factor that reduces the efficiency of antibiotics due to reduced surface area of action, and hence we were keen to take advantage of this synergistic effect.
3.1 Soldiers need shields...maybe - Multidrug Efflux Pump (Bcr)
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