Sampling the cultivation parameter space for the bacterial production of TLR1 intracellular domain reveals the multiple optima
Sampling the cultivation parameter space for the bacterial production of TLR1 intracellular domain reveals the multiple optima
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DOI：
10.1016/j.pep.2021.105832
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发表时间：
2021-02-06
影响因子：
1.6
通讯作者：
Mineev, Konstantin S.
中科院分区：
文献类型：
--
作者：
Goncharuk, Marina V.;Lushpa, Vladislav A.;Mineev, Konstantin S.
T7表达系统是大肠杆菌中生产重组蛋白的一种非常流行的方法，用于结构和功能研究以及治疗应用。有许多有用的工具和成功的技术，允许在这个系统中表达所需的蛋白质。然而，可溶性蛋白质的高产量通常需要系统地优化宽范围的细胞培养参数。在这里，我们分析了三个关键的培养参数-化学诱导剂，温度和诱导后培养时间对可溶性形式的TLR 1胞内TIR结构域表达水平的影响。此外，还研究了Triton X-100去污剂对细胞裂解过程中蛋白溶解度的影响。我们表明，正确折叠的可溶性蛋白质的高表达水平，可以得到不同的组合下的培养参数。
T7 expression system is an extremely popular approach for the recombinant protein production in Escherichia coli for structural and functional studies and therapeutic applications. There are many useful tools and successful techniques that allow expressing the desired protein in this system. However, high yield of soluble protein often requires a systematic optimization of a wide range of cell cultivation parameters. Here we analyze the effect of three key cultivation parameters - chemical inductor, temperature and time of post-induction culturing on the expression level of TLR1 intracellular TIR domain in a soluble form. In addition, the influence of Triton X-100 detergent on the protein solubility during the cell lysis was investigated. We show that a high expression level of the correctly folded soluble protein can be obtained under different combinations of cultivation parameters.