Tungekar Aatir A, Castillo-Corujo Angel, Ruddock Lloyd W
Protein and Structural Biology Research Unit, Faculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu 90220, Finland.
Essays Biochem. 2021 Jul 26;65(2):247-260. doi: 10.1042/EBC20200170.
Recombinant proteins have been extensively employed as therapeutics for the treatment of various critical and life-threatening diseases and as industrial enzymes in high-value industrial processes. Advances in genetic engineering and synthetic biology have broadened the horizon of heterologous protein production using multiple expression platforms. Selection of a suitable expression system depends on a variety of factors ranging from the physicochemical properties of the target protein to economic considerations. For more than 40 years, Escherichia coli has been an established organism of choice for protein production. This review aims to provide a stepwise approach for any researcher embarking on the journey of recombinant protein production in E. coli. We present an overview of the challenges associated with heterologous protein expression, fundamental considerations connected to the protein of interest (POI) and designing expression constructs, as well as insights into recently developed technologies that have contributed to this ever-growing field.
重组蛋白已被广泛用作治疗各种严重和危及生命疾病的疗法，以及高价值工业过程中的工业酶。基因工程和合成生物学的进展拓宽了使用多种表达平台进行异源蛋白生产的视野。选择合适的表达系统取决于多种因素，从目标蛋白的物理化学性质到经济考量。四十多年来，大肠杆菌一直是蛋白质生产的既定选择生物。本综述旨在为任何开始在大肠杆菌中进行重组蛋白生产之旅的研究人员提供一种循序渐进的方法。我们概述了与异源蛋白表达相关的挑战、与目标蛋白（POI）相关的基本考量以及设计表达构建体，还深入探讨了最近为这个不断发展的领域做出贡献的技术。
