Abstract

To increase the production yield of functional recombinant streptavidin in Escherichia coli, the effects of host strains and culture conditions on expression of streptavidin with native C terminal (CNSA, amino acid residues 13 to 159) were investigated. Results show that the CNSA, encoded by the CNSA gene, was produced by E. coliBL21(DE3)pLysS strain in the inclusion body with a high yield up to 46.3% of the total cell protein (about 230 mg/g dry cell weight) after culture condition optimization. The dialysis method was adapted to refold CNSA and the refolding conditions were optimized. More than 90% of inclusion body protein was refolded to mature CNSA under optimized refolding conditions. The purity of the recombinant CNSA achieved 95.0% without using any affinity separation method. Enzyme linked immunosorbent assay (ELISA) analysis indicated that the biotin binding capability of our recombinant CNSA was similar to that of commercial products.

Key words: Streptavidin, Escherichia coli, protein refolding, recombinant protein.