Fusion tags for protein solubility, purification and immunogenicity in Escherichia coli: the novel Fh8 system.
Fusion tags for protein solubility, purification and immunogenicity in Escherichia coli: the novel Fh8 system.
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DOI：
10.3389/fmicb.2014.00063
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发表时间：
2014
影响因子：
5.2
通讯作者：
Domingues L
中科院分区：
文献类型：
--
作者：
Costa S;Almeida A;Castro A;Domingues L
蛋白质现在在各种微生物细胞工厂中广泛生产。大肠杆菌仍然是重组蛋白生产的主要宿主，但作为细菌细胞，它也有其问题：外源蛋白聚集成不溶性包涵体可能是大肠杆菌的主要限制因素。coli表达系统。相反，E.大肠杆菌在成本、易用性和规模方面的优势使得设计新的方法来改善该宿主细胞中的重组蛋白生产是必要的。借助遗传和蛋白质工程，可以设计新的定制策略以满足用户或工艺要求。基因融合技术已被广泛应用于提高大肠杆菌可溶性蛋白的产量和/或纯化。大肠杆菌中表达，并提高了肽的免疫原性。新的融合伴侣不断出现，并补充了传统的解决方案，例如，最近研究的Fh 8融合标签，并被列为最佳溶解度增强剂伴侣。在这篇综述中，我们概述了当前提高大肠杆菌重组蛋白产量的策略。大肠杆菌，包括成功的蛋白质生产的关键因素，突出可溶性蛋白质的生产，并全面总结了最新的和传统上使用的基因融合技术。特别强调了最近发现的Fh 8融合系统，可用于可溶性蛋白的生产，纯化和免疫原性在E。杆菌现有的融合标签的数量可能会在未来几年内增加，应该努力更好地了解融合标签在E.杆菌这些知识无疑将推动开发新的定制工具，用于在这种细菌系统中生产蛋白质。
Proteins are now widely produced in diverse microbial cell factories. The Escherichia coli is still the dominant host for recombinant protein production but, as a bacterial cell, it also has its issues: the aggregation of foreign proteins into insoluble inclusion bodies is perhaps the main limiting factor of the E. coli expression system. Conversely, E. coli benefits of cost, ease of use and scale make it essential to design new approaches directed for improved recombinant protein production in this host cell. With the aid of genetic and protein engineering novel tailored-made strategies can be designed to suit user or process requirements. Gene fusion technology has been widely used for the improvement of soluble protein production and/or purification in E. coli, and for increasing peptide’s immunogenicity as well. New fusion partners are constantly emerging and complementing the traditional solutions, as for instance, the Fh8 fusion tag that has been recently studied and ranked among the best solubility enhancer partners. In this review, we provide an overview of current strategies to improve recombinant protein production in E. coli, including the key factors for successful protein production, highlighting soluble protein production, and a comprehensive summary of the latest available and traditionally used gene fusion technologies. A special emphasis is given to the recently discovered Fh8 fusion system that can be used for soluble protein production, purification, and immunogenicity in E. coli. The number of existing fusion tags will probably increase in the next few years, and efforts should be taken to better understand how fusion tags act in E. coli. This knowledge will undoubtedly drive the development of new tailored-made tools for protein production in this bacterial system.