Zelasko Susan, Palaria Amrita, Das Aditi
Department of Comparative Biosciences, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA.
Protein Expr Purif. 2013 Nov;92(1):77-87. doi: 10.1016/j.pep.2013.07.017. Epub 2013 Aug 20.
Recombinant expression of membrane-bound cytochrome P450s in bacterial expression systems provide a well-established system capable of producing large yields of catalytically active protein. As the biochemical knowledge regarding cytochrome P450s increases, so does the efficiency of protein expression through various modifications that do not disrupt the functional properties of the protein of interest. Changes such as N-terminal modifications, reduction of secondary mRNA structure, bacterial codon usage, selection of vector and host strain, as well as varying external growth conditions all appear to influence protein expression. Several optimizations are often required for sufficient expression of cytochrome P450s at the desired cellular localization. This review aims to comprehensively summarize and update the significant advances made in membrane protein P450 expression in bacterial expression systems.
在细菌表达系统中膜结合细胞色素P450的重组表达提供了一个成熟的系统，能够大量生产具有催化活性的蛋白质。随着关于细胞色素P450的生化知识不断增加，通过各种不破坏目标蛋白质功能特性的修饰来提高蛋白质表达效率的方法也在不断发展。诸如N端修饰、减少二级mRNA结构、细菌密码子使用、载体和宿主菌株的选择以及改变外部生长条件等变化似乎都会影响蛋白质表达。为了使细胞色素P450在所需的细胞定位上充分表达，通常需要进行几种优化。本综述旨在全面总结和更新细菌表达系统中膜蛋白P450表达方面取得的重大进展。
