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Protein Refolding by N-Alkylpyridinium and N-Alkyl-N-methylpyrrolidinium Ionic Liquids

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Abstract

An important property of ionic liquids consisting of cations and anions is that the chemical structures can be easily tuned. To expand the repertoire of effective ionic liquid-based refolding additives, we focused on this tunable property and investigated the effects of new candidates such as N-alkylpyridinium chlorides and N-alkyl-N-methylpyrrolidinium chlorides on protein refolding. Denatured lysozyme (30 mg/mL) was used as a model protein and refolded by 30-fold dilution with various refolding buffers containing different ionic liquids consisting of a systematic variety of alkyl chains. Compared with the refolding yield without additives (lower than 10%), less hydrophobic ionic liquids such as N-ethyl, N-butyl and N-hexylpyridinium chlorides, and N-butyl-N-methylpyrrolidinium chloride were effective in enhancing the refolding yields (46–69%), because they primarily suppressed aggregation because of their chaotropic properties. N-alkylpyridinium cations were more hydrophobic than N-alkyl-N-methylpyrrolidinium cations according to the calculated log P values and prevented aggregation at lower concentrations because of their hydrophobicity. The results provide a range of new effective ionic liquid-based additives for higher protein refolding yields and the knowledge of the effect of chemical structures of additives on protein refolding.

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Acknowledgments

We thank Prof. Kouhei Tsumoto and Dr. Motonori Kudou for providing us with the opportunity to do DSC measurements. This work was supported in part by a Grant-in-Aid for Young Scientists (B) from the MEXT of Japan.

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Authors and Affiliations

  1. Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8656, Japan

    Etsushi Yamamoto, Satoshi Yamaguchi & Teruyuki Nagamune

  2. Department of Bioengineering, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8656, Japan

    Teruyuki Nagamune

  3. Center for NanoBio Integration (CNBI), The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8656, Japan

    Teruyuki Nagamune

Authors
  1. Etsushi Yamamoto
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  2. Satoshi Yamaguchi
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  3. Teruyuki Nagamune
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Correspondence to Satoshi Yamaguchi or Teruyuki Nagamune.

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Yamamoto, E., Yamaguchi, S. & Nagamune, T. Protein Refolding by N-Alkylpyridinium and N-Alkyl-N-methylpyrrolidinium Ionic Liquids. Appl Biochem Biotechnol 164, 957–967 (2011). https://doi.org/10.1007/s12010-011-9187-1

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  • DOI: https://doi.org/10.1007/s12010-011-9187-1

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