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

• Published in: Microbial cell factories Vol. 20; no. 1; p. 208
• Main Authors: Mital, Suraj, Christie, Graham, Dikicioglu, Duygu
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 30-10-2021; BioMed Central; BMC
• Subjects: Bioinformatics; Biotechnology - methods; Biotechnology industry; Chemical engineering; Chemical engineering research; Costs; Design of experiments; Difficult to express enzymes; E coli; Economic analysis; Enzymes; Escherichia coli; Escherichia coli - enzymology; Escherichia coli - genetics; Escherichia coli - metabolism; Experimental design; Gene Expression; Genetic aspects; Grey literature; Immobilized enzymes; Inclusion bodies; Inclusion Bodies - metabolism; Industrial Microbiology; Industrial microorganisms; Manufacturing; Medical research; Microbial enzymes; Microbial genetic engineering; Physiological aspects; Production methods; Production processes; Proteins; Recombinant industrial enzymes; Recombinant Proteins - biosynthesis; Research Design; Review; Solubility; Systematic review; Systems biology; United Kingdom; Systems biology; Escherichia coli; Inclusion bodies; Difficult to express enzymes; Solubility; Recombinant industrial enzymes
• ISSN: 1475-2859; 1475-2859
• 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.