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Approaches for the generation of active papain-like cysteine proteases from inclusion bodies of Escherichia coli

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Abstract

Papain-like cysteine proteases are widely expressed, fulfill specific functions in extracellular matrix turnover, antigen presentation and processing events, and may represent viable drug targets for major diseases. In depth and rigorous studies of the potential for these proteins to be targets for drug development require sufficient amounts of protease protein that can be used for both experimental and therapeutic purposes. Escherichia coli was widely used to express papain-like cysteine proteases, but most of those proteases are produced in insoluble inclusion bodies that need solubilizing, refolding, purifying and activating. Refolding is the most critical step in the process of generating active cysteine proteases and the current approaches to refolding include dialysis, dilution and chromatography. Purification is mainly achieved by various column chromatography. Finally, the attained refolded proteases are examined regarding their protease structures and activities.

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Acknowledgments

This work was supported by Great Project (2011ZX08011-005) from the Major Program of National Science and Technology of China and the National Natural Science Foundation of China (81373128).

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  1. School of Life Science, South China Normal University, 55# Zhongshan Road West, Tianhe District, Guangzhou, 510631, People’s Republic of China

    Chunfang Ling & Deqiu Lin

  2. Guangdong Provincial Key Laboratory of Allergy & Clinical Immunology, The State Key Laboratory of Respiratory Disease, The Second Affiliated Hospital of Guangzhou Medical University, 250# Changgang Road East, Guangzhou, 510260, Guangdong Province, People’s Republic of China

    Chunfang Ling, Junyan Zhang, Deqiu Lin & Ailin Tao

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  1. Chunfang Ling
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Correspondence to Deqiu Lin or Ailin Tao.

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Ling, C., Zhang, J., Lin, D. et al. Approaches for the generation of active papain-like cysteine proteases from inclusion bodies of Escherichia coli . World J Microbiol Biotechnol 31, 681–690 (2015). https://doi.org/10.1007/s11274-015-1804-7

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