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Review
. 2021 Nov;118(11):4159-4167.
doi: 10.1002/bit.27912. Epub 2021 Aug 14.

In vivo cleavage of solubility tags as a tool to enhance the levels of soluble recombinant proteins in Escherichia coli

Filipe S R Silva  1 , Sara P O Santos  1 , Roberto Meyer  1   2 , Eduardo S Silva  1   2 , Carina S Pinheiro  1   2 , Neuza M Alcantara-Neves  1   2 , Luis G C Pacheco  1   3
Affiliations
Review

In vivo cleavage of solubility tags as a tool to enhance the levels of soluble recombinant proteins in Escherichia coli

Filipe S R Silva et al. Biotechnol Bioeng. 2021 Nov.

Abstract

Recombinant proteins are generally fused with solubility enhancer tags to improve the folding and solubility of the target protein of interest. However, the fusion protein strategy usually requires expensive proteases to perform in vitro proteolysis and additional chromatographic steps to obtain tag-free recombinant proteins. Expression systems based on intracellular processing of solubility tags in Escherichia coli, through co-expression of a site-specific protease, simplify the recombinant protein purification process, and promote the screening of molecules that fail to remain soluble after tag removal. High yields of soluble target proteins have already been achieved using these protease co-expression systems. Herein, we review approaches for controlled intracellular processing systems tailored to produce soluble untagged proteins in E. coli. We discuss the different genetic systems available for intracellular processing of recombinant proteins regarding system design features, advantages, and limitations of the various strategies.

Keywords: Escherichia coli; controlled intracellular processing; protein solubility; recombinant proteins; site-specific protease.

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