Shafiee Fatemeh, Sharifi Shima, Amini Armin
Department of Pharmaceutical Biotechnology, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran.
Bioinformatics Research Center, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran.
Iran J Biotechnol. 2024 Apr 1;22(2):e3728. doi: 10.30498/ijb.2024.400460.3728. eCollection 2024 Apr.
The main problem in the recombinant protein expression in strains, especially for high-yield production, is the accumulation in un-folded and inactive inclusion bodies. A suitable solution is the direction into the soluble cytoplasmic products by solubilizing tags. The use of inteins with self-cleaving ability, in addition to increase the chance of soluble protein expression, facilitates their purification process.
In this review article, papers related to the use of intein tags for soluble expression or protein purification were collected regardless the time limit. Available databases including Pubmed, google scholar, ScienceDirect, Web of Science, Scopus, and Embase was searched. The best condition for soluble expression or purification was focused in all articles.
There are various intein tags commercially available in expression vectors that results in gaining our goal in facilitating the recombinant protein solubilization as well as its simple purification. It is enough to induce the self-cleavage property of the intein, which varies according to the type of intein used. In this way, the target protein is easily separated from the purification tag without the need to add protease enzymes such as enterokinase or treatment with various chemicals. The most common affinity tag in intein-based systems is Chitin Binding Domain attached to the chitin resin.
In this review article, we introduced proteins or peptides which produced in fusion to intein tags and discussed about their expression condition and purification process in order to enhance the chance of soluble expression and intein cleavage in a single stage, respectively.
在菌株中进行重组蛋白表达时，主要问题，尤其是对于高产生产而言，是未折叠且无活性的包涵体的积累。一种合适的解决方案是通过可溶标签将其导向可溶性细胞质产物。使用具有自我切割能力的内含肽，除了增加可溶性蛋白表达的机会外，还便于其纯化过程。
在这篇综述文章中，收集了与使用内含肽标签进行可溶性表达或蛋白纯化相关的论文，不设时间限制。搜索了包括PubMed、谷歌学术、ScienceDirect、科学网、Scopus和Embase在内的可用数据库。所有文章都聚焦于可溶性表达或纯化的最佳条件。
表达载体中有多种市售的内含肽标签，这有助于实现我们促进重组蛋白溶解及其简单纯化的目标。诱导内含肽的自我切割特性就足够了，其会根据所使用的内含肽类型而有所不同。通过这种方式，无需添加诸如肠激酶等蛋白酶或用各种化学物质处理，目标蛋白就能轻松地与纯化标签分离。基于内含肽的系统中最常见的亲和标签是附着在几丁质树脂上的几丁质结合域。
在这篇综述文章中，我们介绍了与内含肽标签融合产生的蛋白质或肽，并分别讨论了它们的表达条件和纯化过程，以提高可溶性表达的机会和在单一阶段进行内含肽切割的效率。
