Optimization of culture media for extracellular expression of streptokinase in Escherichia coli using response surface methodology in combination with Plackett-Burman Design

Purpose: To investigate the enhancement of streptokinase extracellular expression in Escherichia coli by adjusting culture media. Methods: Screening of 10 chemical factors (EDTA, peptone, glycine, triton X-100, glycerol, K 2 HPO 4 , KH 2 PO 4 , Ca 2+ (calcium chloride), yeast and NaCl) in order to increase the secretion of extracellular protein was carried out by response surface methodology (RSM). The method was also employed to optimize the concentrations of critical factors that had been determined in the screening step. Results : The results indicate that glycine, triton X-100 and Ca 2+ were the most effective chemical factors in terms of increase in extracellular expression of streptokinase with optimum levels of 0.878, 0.479 and 0.222 %, respectively. Expression of streptokinase under optimum concentrations of critical permeabilizing factors led to a 7-fold increase in the quantity of secreted recombinant protein (5824 U/mL) compared to the initial level (802 U/mL). Conclusion : The results show that medium optimization using RSM is effective in improving extracellular streptokinase expression. The optimization medium is considered fundamental and useful for efficient production of streptokinase on a large scale. Keywords : Streptokinase, Response surface methodology, Membrane permeabilization, Extracellular secretion