Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

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

Mojtaba Aghaeepoor^1,2, Farzad Kobarfard³, Mohammad Reza Akbari Eidgahi¹, Aliakbar Shabani¹, Ehsan Dehnavi², Maryam Ahmadi¹, Ali Akbarzadeh²

¹Semnan Biotechnology Research Center, Semnan University of Medical Sciences, Basij Blvd, Semnan; ²Gene Transfer Pioneers (GTP) Research Group; ³Department of Medicinal Chemistry, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Vali Asr Ave, Tehran, Iran.

For correspondence:- Ali Akbarzadeh Email: akbarzadeh80@yahoo.com Tel:+982188200114

Accepted: 15 October 2017 Published: 30 November 2017

Citation: Aghaeepoor M, Kobarfard F, Eidgahi MR, Shabani A, Dehnavi E, Ahmadi M, et al. Optimization of culture media for extracellular expression of streptokinase in Escherichia coli using response surface methodology in combination with Plackett-Burman Design. Trop J Pharm Res 2017; 16(11):2567-2576 doi: 10.4314/tjpr.v16i11.1

© 2017 The authors.
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Abstract

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, K₂HPO₄, KH₂PO₄, Ca²⁺ (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 Ca²⁺ 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