loading page

In vivo cleavage of solubility tags as a tool to enhance the levels of soluble recombinant proteins in Escherichia coli
  • +4
  • Filipe Silva,
  • Sara Santos,
  • Roberto Meyer,
  • Eduardo Silva,
  • Carina Pinheiro,
  • Neuza Alcantara-Neves,
  • Luis Pacheco
Filipe Silva
Federal University of Bahia

Corresponding Author:[email protected]

Author Profile
Sara Santos
Federal University of Bahia
Author Profile
Roberto Meyer
Federal University of Bahia
Author Profile
Eduardo Silva
Federal University of Bahia
Author Profile
Carina Pinheiro
Federal University of Bahia
Author Profile
Neuza Alcantara-Neves
Federal University of Bahia
Author Profile
Luis Pacheco
Federal University of Bahia
Author Profile

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.
03 Feb 2021Submitted to Biotechnology and Bioengineering
03 Feb 2021Submission Checks Completed
03 Feb 2021Assigned to Editor
24 Feb 2021Reviewer(s) Assigned
10 Jun 2021Editorial Decision: Revise Major
10 Jun 2021Review(s) Completed, Editorial Evaluation Pending
02 Jul 20211st Revision Received
03 Jul 2021Submission Checks Completed
03 Jul 2021Assigned to Editor
12 Jul 2021Reviewer(s) Assigned
06 Aug 2021Editorial Decision: Accept
06 Aug 2021Review(s) Completed, Editorial Evaluation Pending
Nov 2021Published in Biotechnology and Bioengineering volume 118 issue 11 on pages 4159-4167. 10.1002/bit.27912