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

Kuniaki Nagamine, Shiho Onodera, Ai Kurihara, Tomoyuki Yasukawa, Hitoshi Shiku, Ryutaro Asano, Izumi Kumagai, Tomokazu Matsue

Research output: Contribution to journalArticlepeer-review

15 Citations (Scopus)

Abstract

A microbial array chip with collagen gel spots entrapping living Escherichia coli (E. coli) DH5α was applied for the screening of recombinant protein solubilities. The α-fragment of β-galactosidase (βGal) 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 βGal. 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 βGal activity and therefore, with the soluble fraction of MBP in the host cells.

Original languageEnglish
Pages (from-to)1008-1013
Number of pages6
JournalBiotechnology and Bioengineering
Volume96
Issue number5
DOIs
Publication statusPublished - 2007 Apr 1

Keywords

  • High-throughput
  • Microbial array chip
  • Proteome
  • Recombinant protein solubility
  • Scanning electrochemical microscopy

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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