Strategies for optimization of heterologous protein expression in E-coli: Roadblocks and reinforcements
Strategies for optimization of heterologous protein expression in E-coli: Roadblocks and reinforcements
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DOI：
10.1016/j.ijbiomac.2017.08.080
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发表时间：
2018-01-01
影响因子：
8.2
通讯作者：
Kaur, Jagdeep
中科院分区：
文献类型：
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作者：
Kaur, Jashandeep;Kumar, Arbind;Kaur, Jagdeep
大肠杆菌是在细菌中生产重组蛋白的最首选的系统，改进的遗传工具/方法的可用性使其比以往任何时候都更有价值。这种表达系统面临的主要挑战是表达具有稀有密码子或膜的异常困难/复杂的蛋白和有毒蛋白。自然界中大量表达或疏水性表达的蛋白质往往会形成不溶于水的团块。尽管有适当的表达系统，但一些蛋白质的表达水平很低，甚至根本不表达。选择正确的表达系统/方案对于以天然形式大量表达蛋白质是必不可少的。一些载体、它们的相容宿主和培养条件可用于大量和以天然形式表达重组蛋白。此外，带有融合标签/伴侣的载体除了恢复蛋白质的天然结构外，还促进了蛋白质在可溶性部分的表达，并有助于蛋白质的正确折叠。通过优化复性条件，可以从难溶的包涵体中回收天然蛋白质。本文综述了在大肠杆菌中成功表达外源基因的原核表达系统的最新进展，并重点介绍了提高天然重组蛋白产量的策略。(C)2017爱思唯尔B.V.保留所有权利。
E. coli is most preferred system used for the production of recombinant proteins in bacteria and the availability of improved genetic tools/methods are making it more valuable than ever. Major challenges faced by this expression system are the expression of unusually difficult/complex proteins with rare codons or membrane and toxic proteins. The proteins expressed either in large amount or hydrophobic in nature tend to form insoluble mass. Despite the appropriate expression system, some proteins express at very low level or not at all. Choosing the correct expression system/protocols are obligatory for the substantial expression of protein in the native form. A number of vectors, their compatible hosts and culture conditions can be used to express recombinant proteins in large amounts and in native form. Also, vectors with the fusion tags/chaperons facilitate protein expression in soluble fraction and assist in proper protein folding besides restoring the native structure of protein. The recovery of native proteins from insoluble inclusion bodies can be achieved by optimization of refolding conditions. In the present review, we discussed recent updates on prokaryotic expression system for successful heterologous gene expression in E. coli and focused on strategies to maximize the yields of native recombinant proteins. (C) 2017 Elsevier B.V. All rights reserved.