Aghaeepoor Mojtaba, Akbarzadeh Ali, Kobarfard Farzad, Shabani Ali Akbar, Dehnavi Ehsan, Jamshidi Aval Sanaz, Akbari Eidgahi Mohammad Reza
Department and Biotechnology Research Center, Semnan University of Medical Sciences, Semnan, Iran.
Student Research Committee, Semnan University of Medical Sciences, Semnan, Iran.
Iran J Pharm Res. 2019 Spring;18(2):961-973. doi: 10.22037/ijpr.2019.1100636.
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 investigation. 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 (OD). 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 (OD). These settings result in 4.14fold 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.
链激酶（SK）是一种由414个氨基酸组成的细胞外蛋白，作为一种常用的溶栓剂具有相当重要的临床意义。由于其广泛的应用和临床重要性，设计更高效的SK生产平台值得研究。在这方面，一个合成的SK基因被优化并克隆到pET21b质粒中用于周质表达。采用响应面法设计了总共20个实验，以优化异丙基-β-D-硫代半乳糖苷（IPTG）浓度、诱导后时间和诱导时的细胞密度（OD）。选定参数的最佳水平成功确定为：IPTG浓度为0.28 mM，诱导后时间为9.889小时，细胞密度（OD）为3.40768。与初始表达条件（1853 IU/mL）相比，这些设置使最佳表达条件下（7663 IU/mL）的SK生产率提高了4.14倍。在摇瓶中实现更高的SK产量可导致该药物更具成本效益的工业化生产，这是SK生产研究的最终目标。
