Department of Chemical Engineering and Biotechnology, University of Cambridge, Cambridge, CB3 0AS, UK.
Department of Biochemical Engineering, University College London, London, WC1E 6BT, UK.
Microb Cell Fact. 2021 Oct 30;20(1):208. doi: 10.1186/s12934-021-01698-w.
Recombinant enzyme expression in Escherichia coli is one of the most popular methods to produce bulk concentrations of protein product. However, this method is often limited by the inadvertent formation of inclusion bodies. Our analysis systematically reviews literature from 2010 to 2021 and details the methods and strategies researchers have utilized for expression of difficult to express (DtE), industrially relevant recombinant enzymes in E. coli expression strains. Our review identifies an absence of a coherent strategy with disparate practices being used to promote solubility. We discuss the potential to approach recombinant expression systematically, with the aid of modern bioinformatics, modelling, and 'omics' based systems-level analysis techniques to provide a structured, holistic approach. Our analysis also identifies potential gaps in the methods used to report metadata in publications and the impact on the reproducibility and growth of the research in this field.
在大肠杆菌中重组酶的表达是生产大量蛋白质产品最常用的方法之一。然而，这种方法常常受到不希望的包涵体形成的限制。我们的分析系统地回顾了 2010 年至 2021 年的文献，并详细介绍了研究人员在大肠杆菌表达菌株中表达困难表达（DtE）、工业相关重组酶所采用的方法和策略。我们的综述发现，缺乏一致的策略，不同的方法被用于促进可溶性。我们讨论了在现代生物信息学、建模和基于“组学”的系统级分析技术的帮助下，系统地处理重组表达的可能性，为提供一种结构化、整体的方法。我们的分析还确定了在报告出版物元数据的方法中存在的潜在差距，以及对该领域研究的可重复性和发展的影响。
