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Expression in Bacteria and Refolding

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Advanced Methods in Structural Biology

Part of the book series: Springer Protocols Handbooks ((SPH))

Abstract

Production of proteins by bacterial expression is common due to straightforwardness and inexpensiveness. In this chapter, we focus on the expression system using Escherichia coli and refolding of inclusion bodies formed with insoluble protein materials. In the first half, several steps required to produce soluble proteins as much amount as possible, in E. coli, are described. Here, the choice of either vector or bacterial strains, induction, extraction, fusion of solubilizing tags, and strategies to facilitate disulfide bond formation are included. In the second half, strategies to get soluble proteins from inclusion bodies are described. Here, the mechanism of protein refolding, the isolation of inclusion bodies, the choice of solubilizing materials, the refolding step, and the effects of additives while refolding are included. The selection of various strategies in these steps is discussed.

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Authors and Affiliations

  1. Department of Bioengineering, School of Engineering, The University of Tokyo, Tokyo, Japan

    Hiroki Akiba & Kouhei Tsumoto

  2. Medical Proteomics Laboratory, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan

    Kouhei Tsumoto

Authors
  1. Hiroki Akiba
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  2. Kouhei Tsumoto
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Correspondence to Kouhei Tsumoto .

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Editors and Affiliations

  1. Institute of Materials Structure Science, High Energy Accelerator Research Organization (KEK) , Tsukuba, Ibaraki, Japan

    Toshiya Senda

  2. Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Hokkaido, Japan

    Katsumi Maenaka

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© 2016 Springer Japan

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Akiba, H., Tsumoto, K. (2016). Expression in Bacteria and Refolding. In: Senda, T., Maenaka, K. (eds) Advanced Methods in Structural Biology. Springer Protocols Handbooks. Springer, Tokyo. https://doi.org/10.1007/978-4-431-56030-2_1

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  • DOI: https://doi.org/10.1007/978-4-431-56030-2_1

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  • Print ISBN: 978-4-431-56028-9

  • Online ISBN: 978-4-431-56030-2

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