Optimization of Expression and Purification Procedure of Recombinant Human Collagen Peptide

To optimize the fermentation and purification conditions of recombinant human collagen peptide(CP6) in E.coli pC6-BL21,the temperature,opportunity and time for induction as well as inducer concentration for CP6 expression in recombinant E.coli was optimized.The optimal induction and expression conditions were as follows,after inoculating at an amount of 2%,the bacteria were fermented for 3.0 h at 37 ℃;then 0.5 mmol/L IPTG was added as inducer;and the peptide was expressed under 30 ℃ for 5.0 h.After fermented under the optimal conditions,the lysate of bacteria was purified by nickel ion affinity;and purity of target protein was determined by SDS-PAGE.Results showed that pure protein could be obtained if eluted by 150 mmol/L imidazole buffer after being purified,and western blotting showed that the expressed protein could specifically bind with human monoclonal antibody COL6A2.