Imperial College London/M2/Detail

E.coli is naturally equipped with a multitude of defences to enable colonisation of the intestine. We are using two global transcription factors (RcsB & YgiV) to hijack this natural process in a way that maximises acid resitance. We have additionally upregulated a third enzyme (rfal) to enhance the encapsulation of single cells (over and above colony encapsulation). Finally, the two biosynthetic genes (OtsA & OtsB) code for the production of trehalose. Our manipulation of endogenous pathways reduces virulence while enchancing pill functionality.

Colanic acid encapsulation and the synthesis of various acid resistance proteins protect the protein of interest from the digestive assaults of the buccal cavity and acid-filled stomach. Once the pill reaches the intestine, gut microflora will digest the E.ncapsulator's colanic acid coat, releasing the protein of interest.

Trehalose preserves our protein of interest in its correct conformation in response to dessication. Thus trehalose allows for the freeze drying of the pill, and this will allow easy transport and storage.

Encapsulation is initiated following the completion of protein production (Module 1). It should be noted that the protein production of our compound of interest continues at a lower 'maintenance level' throughout Module 2.