Role of molecular and chemical chaperon in assisting refolding of BMP2 in E. coli

Bone morphogenetic protein 2 (BMP2) when expressed in bacteria forms inclusion bodies (IBs) due to its complex disulfide-rich structure. Chaperons are already well known for their role in assisting protein folding. In our studies, we have used two E. coli strains, BL21(DE3) and SHuffle® T7 cells for overexpressing BMP2 in soluble fraction. We observed that SHuffle® T7 cells successfully expressed soluble functionally active BMP2 in presence of molecular and chemical chaperones at low temperature. The combination of chemical and molecular chaperons further increases the yield of protein. The best-suited chaperon system for overexpression of BMP2 is GroES-GroEL at low temperature. The soluble functionally active BMP2 is confirmed by its binding to its receptor ALK3 through Native PAGE and ELISA assay using BMP2 specific antibody. It is possible to obtain BMP2 expression in soluble active form by using molecular and chemical chaperons which work synergistically in bacterial cells to fold disulphide-rich proteins at low temperature in easy and time saving steps (18 ̊C).