Electrochemical screening of recombinant protein solubility in Escherichia coli using scanning electrochemical microscopy (SECM) |
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Authors: | Nagamine Kuniaki Onodera Shiho Kurihara Ai Yasukawa Tomoyuki Shiku Hitoshi Asano Ryutaro Kumagai Izumi Matsue Tomokazu |
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Institution: | Graduate School of Environmental Studies, Tohoku University, 6-6-11 Aramaki Aoba, Aoba-ku, Sendai 980-8579, Japan. |
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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. |
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Keywords: | microbial array chip recombinant protein solubility scanning electrochemical microscopy high‐throughput proteome |
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