Electrochemical screening of recombinant protein solubility in Escherichia coli using scanning electrochemical microscopy (SECM)

Biotechnol Bioeng. 2007 Apr 1;96(5):1008-13. doi: 10.1002/bit.21173.

Abstract

A microbial array chip with collagen gel spots entrapping living Escherichia coli (E. coli) DH5alpha was applied for the screening of recombinant protein solubilities. The alpha-fragment of beta-galactosidase (betaGal) was fused to the target protein, namely, maltose-binding protein (MBP), to monitor the solubility of MBP. Scanning electrochemical microscopy (SECM) was used to detect the release of p-aminophenol from E. coli cells catalyzed by intracellular betaGal. Comparison of the SECM-based method with the Western blotting-based method indicated that the current response obtained using SECM increased with an increase in the betaGal activity and therefore, with the soluble fraction of MBP in the host cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carrier Proteins / chemistry*
  • Carrier Proteins / metabolism
  • Electrochemistry / methods*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Maltose-Binding Proteins
  • Microscopy / methods*
  • Microscopy, Electron, Scanning
  • Recombinant Proteins / chemistry*
  • Recombinant Proteins / metabolism

Substances

  • Carrier Proteins
  • Maltose-Binding Proteins
  • Recombinant Proteins