Module 2 - Encapsulsation

Overview

Module 2: Encapsulation

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.

By 'hacking' E.coli’s endogenous acid resistance pathways in three places we induced 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. RcsB up regulation = Induces colanic acid biosynthesis.

2. YgiV up regulation = Blocks biofilm formation and enhanced colanic acid production.

3. Rfal up regulation = Tethers colanic acid to the cell surface creating a microcapsule.

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.