Module 2 - Encapsulsation

Overview

Aside from aiding in pill manufacture, encapsulation (Module 2) allows the safe passage of polypetides through the stomach into the intestine. Without encapsulation, our polypeptides would be denatured and degraded by stomach acid and digestive proteases respectively.

We used a 'Triple Hack' to rewire E.coli’s endogenous acid resistnace pathways in three places. As a result, we can induce the formation of a safe colanic acid microcapsule that protects our chassis from the harsh acidic conditions of the stomach.

The Encapsulation Triple Hack:

1. Up regulation of colonic acid biosynthesis with RcsB.

2. Enhancing RcsB activity and blocking biofilm formation with YgiV.

3. Tethering colonic acid to the cell surface with Rfal.

In addition, Module 2 involves the synthesis of the preservative trehalose. This is a disaccharide that allows us to freeze dry the chassis without damaging either its structural integrity or the intracellular polypeptides. Freeze drying is an important storage mechanism that will facilitate the widespread dispersal of E.ncapsulator technology.