Optimization and High Level Production of Recombinant Synthetic Streptokinase in E. coli Using Response Surface Methodology

Document Type: Research article

Authors

1 Department and Biotechnology Research Center, Semnan University of Medical Sciences, Semnan, Iran.

2 Student Research Committee, Semnan University of Medical Sciences, Semnan, Iran.

3 Gene Transfer Pioneers (GTP) Research Group, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

4 Department of Medicinal Chemistry, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Abstract

Streptokinase (SK) is an extracellular protein comprising 414 amino acids with considerable clinical importance as a commonly used thrombolytic agent. Due to its wide spread application and clinical importance designing more efficient SK production platforms worth investigatinginvestigation. In this regard, a synthetic SK gene was optimized and cloned in to pET21b plasmid for periplasmic expression. Response surface methodology was used to design a total of 20 experiments for optimization of IPTG concentration, post-induction period, and cell density of induction (OD600). The optimum levels of the selected parameters were successfully determined to be 0.28 mM for IPTG concentration, 9.889 H for post induction period, and 3.40768 for cell density (OD600). These settings result in 4.14 fold increase in SK production rate of optimum expression conditions (7663 IU/mL) in comparison to the primary expression conditions (1853 IU/mL). Achieving higher yields of SK production in shake flask could lead to more cost effective industrial production of this drug which is the ultimate aim of SK production studies.

Keywords

Main Subjects