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) 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.
authors.affiliation: Graduate School of Environmental Studies, Tohoku University, 6-6-11 Aramaki Aoba, Aoba-ku, Sendai 980-8579, Japan.
authors.forename: Kuniaki
authors.initials: K
authors.lastname: Nagamine
authors.forename: Shiho
authors.initials: S
authors.lastname: Onodera
authors.forename: Ai
authors.initials: A
authors.lastname: Kurihara
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Yasukawa
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Shiku
authors.forename: Ryutaro
authors.initials: R
authors.lastname: Asano
authors.forename: Izumi
authors.initials: I
authors.lastname: Kumagai
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Matsue
mesh_terms: D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D004563:Electrochemistry / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D058619:Maltose-Binding Proteins; D008853:Microscopy / Q000379:methods*; D008855:Microscopy, Electron, Scanning; D011994:Recombinant Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D058619:Maltose-Binding Proteins; D011994:Recombinant Proteins