Recombinant expression of insoluble enzymes in Escherichia coli: a systematic review of experimental design and its manufacturing implications


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Authors
Mital, Suraj 
Christie, Graham 
Abstract

Abstract: 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.

Description

Funder: Johnson Matthey Cambridge

Keywords
Review, Escherichia coli, Recombinant industrial enzymes, Difficult to express enzymes, Inclusion bodies, Systems biology, Solubility
Journal Title
Microbial Cell Factories
Conference Name
Journal ISSN
1475-2859
Volume Title
20
Publisher
BioMed Central