Stability and Biological Activity of Human Intestinal Trefoil Factor Produced by Pichia pastoris

Intestinal trefoil factor (ITF), a member of the trefoil factor family, plays an important role in protecting the epithelial layer of the intestinal tract from damage and repairing epithelium after injury. In the present study, we expressed recombinant hITF by Picha pastoris expression system in shake flasks to 50mg/L. rhITF was purified up to 95% by ammonium sulfate precipitation, Ni-NTA affinity chromatography and ultrafiltration. In conditions simulating intestinal tract environment, rhITF was shown to exhibit resistance to proteases, heat, acid and alkali. In addition, rhITF was found to be biologically active in an in vitro restitution model. This study lays the foundation for the commercial production of rhITF and provides theoretical evidence for the oral use of rhITF.