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

期刊

BIOTECHNOLOGY AND BIOENGINEERING
卷 118, 期 11, 页码 4159-4167

出版社

WILEY
DOI: 10.1002/bit.27912

关键词

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

资金

  1. FAPESB
  2. National Council for Scientific and Technological Development of Brazil (CNPq)
  3. FAPESB/CNPq PRONEM-2014 grant

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The article discusses methods for producing soluble untagged proteins in Escherichia coli using genetic engineering techniques, which involve intracellular processing through co-expression of a specific protease. This approach simplifies the purification of recombinant proteins, improves the solubility of target proteins, and facilitates screening for proteins that remain soluble after tag removal.
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.

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