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

doi:10.1002/bit.21173

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 journal › Article › peer-review

16 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 language	English
Pages (from-to)	1008-1013
Number of pages	6
Journal	Biotechnology and Bioengineering
Volume	96
Issue number	5
DOIs	https://doi.org/10.1002/bit.21173
Publication status	Published - 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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10.1002/bit.21173

Cite this

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

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.",

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

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T1 - Electrochemical screening of recombinant protein solubility in Escherichia coli using scanning electrochemical microscopy (SECM)

AU - Nagamine, Kuniaki

AU - Onodera, Shiho

AU - Kurihara, Ai

AU - Yasukawa, Tomoyuki

AU - Shiku, Hitoshi

AU - Asano, Ryutaro

AU - Kumagai, Izumi

AU - Matsue, Tomokazu

PY - 2007/4/1

Y1 - 2007/4/1

N2 - 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.

AB - 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.

KW - High-throughput

KW - Microbial array chip

KW - Proteome

KW - Recombinant protein solubility

KW - Scanning electrochemical microscopy

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JF - Biotechnology and Bioengineering

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IS - 5

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

Abstract

Keywords

ASJC Scopus subject areas

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