促进包涵体蛋白复性的几种有效添加剂

利用大肠杆菌高水平表达重组蛋白时,由于缺乏翻译后加工过程,目的蛋白通常以无活性的包涵体的形式产生。在包涵体蛋白复性过程中,溶解的蛋白质容易相互聚集生成沉淀,成为复性技术中最难以克服的问题。目前通常的做法是在复性过程中加入适当的添加剂,使其在最佳反应条件下促进蛋白质重折叠为活性形式。本文旨在介绍几种有效的促进复性的添加剂及其最佳使用条件,为研究者提供一定的借鉴。

Several Additives That Effectively Improve Protein Refolding

Due to the lack of machineries for post-translational modifications, the use of E. coli for large-scale protein production is frequently plagued by the formation of insoluble protein aggregates , inclusion bodies. Refolding of protein from inclusion bodies were first denatured by using high dpnaturant and detergent. After protein was redissolved in the low concentration denaturant, protein tend to aggregate because of molecular interaction. Protein aggregation is believed to be the main culprit for low yields of recombinant protein. Addition of suitable additives into the refolding buffer can significantly improve protein renaturation under the high protein concentrations. Reviewed in his are several protein refolding promoters, including controlled air oxidation, SDS, PEG, NDSBs, Brij 58P, L-Arginine. The principles and optimal conditions were also discussed.