Optimization of Fermentation Conditions for Reteplase Expression by Escherichia coli Using Response Surface Methodology.

摘要: Background Expression of heterologous proteins at large scale is often a challenging job due to plasmid instability, accumulation acetate and oxidative damage in bioreactors. Therefore, it necessary optimize parameters influencing cell growth expression recombinant protein. Methods In the present study, optimal culture conditions for reteplase by Escherichia coli (E. coli) BL21 (DE3) bench-top bioreactor was determined. Response Surface Methodology (RSM) based on Box-Behnken design used evaluate effect three variables (i.e., temperature, shaking speed pH) their interactions with cellular protein production. The obtained data were analyzed Design Expert software. Results Based results 15 experiments, response surface quadratic model developed which explain relation between production investigated variables. high value "R-Squared" (0.9894) F-value 51.99 confirmed accuracy this model. According model, optimum fermentation temperature 32°C, 210 rpm, pH 8.4. This predicted condition applied leading yield 188 mg/l. Conclusion Our indicate significant role (e.g., pH, oxygen supply) confirm need optimization. proposed strategy here can also be experimental set-up optimization other proteins.