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Refolding of Laccase in Dilution Additive Mode with Copper-Based Ionic Liquid

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Abstract

Ionic liquids (ILs) are molten salts which do not crystallize at room temperature. Tunable physicochemical properties of ILs including hydrophobicity and polarity facilitate their applications in many biological processes. In this study, a copper-based IL was employed in order to enhance the refolding efficiency of laccase from Trametes versicolor which requires copper as a cofactor. When 1-ethyl-3-methylimidazolium trichlorocuprate ([EMIM][CuCl3]) was added to refolding buffer instead of urea, the laccase refolding yield was improved more than 2.7 times compared to the conventional refolding buffer which contains urea. When the refolding of laccase was carried out at different temperatures (4, 25, and 37 °C), the highest refolding yield was obtained at 25 °C. At low temperature, two conflicting effects, i.e., suppression of the aggregate formation and decrease of folding rate, influence the protein refolding. In contrast, a copper-based IL did not enhance the refolding of lysozyme, a non-copper-containing protein. From these results, we can conclude that this copper-based IL, [EMIM][CuCl3], was exclusively effective on the refolding process of a copper-containing protein.

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Acknowledgments

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea funded by the Ministry of Education, Science and Technology (grant number 2010-0013308).

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Authors and Affiliations

  1. Department of Biological Engineering, Inha University, Incheon, 402-751, Republic of Korea

    Sang-Woo Bae, Kihun Ahn & Yoon-Mo Koo

  2. Department of Chemical Engineering and Nano-Bio Technology, Hannam University, Daejeon, 305-811, Republic of Korea

    Sung Ho Ha

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Correspondence to Yoon-Mo Koo or Sung Ho Ha.

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Bae, SW., Ahn, K., Koo, YM. et al. Refolding of Laccase in Dilution Additive Mode with Copper-Based Ionic Liquid. Appl Biochem Biotechnol 171, 1289–1298 (2013). https://doi.org/10.1007/s12010-013-0422-9

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