Improved Production of Recombinant Human Activin A in Escherichia coli

Abstract

Activin A as a member of transforming growth factor β (TGF-β) superfamily, plays important roles in the body such as regulation of cell growth and differentiation, wound repairing and modulation of inflammatory responses. More importantly, it can be used as a therapeutic agent; so its recombinant production, especially in the periplasmic space of E. coli as an economical bacterium is of great value. The aim of this study is large- scale production of activin A with a correct structure. For this purpose, three strategies were used. First, an efficient and appropriate signal peptide, modified Iranian Bacillus Licheniformis α-amylase signal peptide, was used to secrete activin A to the periplasmic space of E. coli as a suitable environment for correct protein folding. Second, cytoplasmic chaperones, Dnak, DnaJ, GroEL/ GroES, TF (trigger factor) were expressed simultaneously with activin A. And finally, the optimal agitation rate for improved production of activin A was found in the bioreactor scale. Our results indicated that by the co-expression of TF with activin A and using agitation rate of 1000 rpm maximum expression of activin A in E. coli was obtained. More importantly, based on the CD spectroscopy results and bioassay test the recombinant activin A had appropriate secondary structure and was fully functional.