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In vivo cleavage of solubility tags as a tool to enhance the levels of soluble recombinant proteins in Escherichia coli
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  • Filipe Silva,
  • Sara Santos,
  • Roberto Meyer,
  • Eduardo Silva,
  • Carina Pinheiro,
  • Neuza Alcantara-Neves,
  • Luis Pacheco
Filipe Silva
Federal University of Bahia
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Sara Santos
Federal University of Bahia
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Roberto Meyer
Federal University of Bahia
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Eduardo Silva
Federal University of Bahia
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Carina Pinheiro
Federal University of Bahia
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Neuza Alcantara-Neves
Federal University of Bahia
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Luis Pacheco
Federal University of Bahia
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Abstract

Recombinant proteins are generally fused with solubility enhancer tags to improve target protein folding and solubility. However, the fusion protein strategy usually requires the use of expensive proteases to perform in vitro proteolysis and additional chromatography 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, are useful for simplifying the recombinant protein purification process, for screening molecules that fail to remain soluble after tag removal, and to promote higher yields of soluble target protein. Herein, we review controlled intracellular processing (CIP) systems, tailored to produce soluble untagged proteins in E. coli. We discuss the different genetic systems available for intracellular protein processing regarding system design features, significant advantages and limitations of the various strategies.

Peer review status:IN REVISION

03 Feb 2021Submitted to Biotechnology and Bioengineering
03 Feb 2021Assigned to Editor
03 Feb 2021Submission Checks Completed
24 Feb 2021Reviewer(s) Assigned
10 Jun 2021Review(s) Completed, Editorial Evaluation Pending
10 Jun 2021Editorial Decision: Revise Major