Abstract
A thorough characterisation of the genetics, physiology and metabolism of Escherichia coli has led to the availability of a large number of strains and vectors suitable for recombinant protein expression. Despite the relative ease in using E. coli for achieving amplified expression of many recombinant proteins, for some proteins this can be a frustrating and time-consuming process leading to very low expression or no expression at all. This is especially true for membrane proteins, which introduce additional challenges. A number of factors can be considered and optimised for achieving required levels of amplified expression of recombinant proteins in E. coli that are broadly classified as host strain, expression vector and growth conditions. In this paper we summarise these factors and consolidate the common challenges encountered and approaches to overcome them, focusing in particular on cases where there is low amplified expression or no expression at all of the desired recombinant protein, due to various reasons.
•Escherichia coli is the most widely used organism for amplifying recombinant protein expression.•This is challenging for some proteins, giving low expression or no expression at all.•Different parameters can be altered for optimising recombinant protein production.•These include the host strain, the expression vector and growth conditions.•Issues at the DNA level and protein toxicity/aggregation/instability can be overcome.