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Production of Bioactive Human Beta-Defensin-4 in Escherichia coli Using SUMO Fusion Partner

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Abstract

The human beta defensins-4 (hBD4) exhibit a broad range of antimicrobial properties and are thought to be ideal therapeutic agents because of their potential ability to circumvent the problems of acquired resistance often observed with other antimicrobial therapies. We report here the application of small ubiquitin-related modifier (SUMO) fusion technology to the expression and purification of cationic antibacterial peptide hBD4. The fusion protein expressed in a soluble form was purified to a purity of 90% by Ni-IDA chromatography and 637 mg protein of interest was obtained per liter of fermentation culture. After the SUMO-hBD4 fusion protein was cleaved by the SUMO protease at 30 °C for 1 h, the cleaved sample was re-applied to a Ni-IDA. Finally, about 166 mg recombinant hBD4 was obtained from 1 L fermentation culture with no less than 96% purity and the recombinant hBD4 had similar antimicrobial properties to the synthetic hBD4. Thus, the SUMO-mediated peptide expression and purification system potentially could be employed for the production of recombinant cytotoxic peptides.

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Abbreviations

hBD4:

The human beta defensins-4

SUMO:

Small ubiquitin-related modifier

IPTG:

Isopropyl-β-D-1-thiogalactopyranoside

MIC:

The minimal inhibitory concentration

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Acknowledgments

This work was financially supported by National Nature Science Foundation of China, (No. 30270193) and Natural Science Foundation of Jiangsu Province, China (No. BK2006221).

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Authors and Affiliations

  1. Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Science College, Nanjing Normal University, 210046, Nanjing, Jiangsu, People’s Republic of China

    Jian Feng Li, Jie Zhang, Zhen Zhang, Hong Wei Ma, Jia Xin Zhang & Shuang Quan Zhang

  2. Jiangsu Engineering Research Center for Biomedical Function Materials, Nanjing Normal University, Nanjing, People’s Republic of China

    Shuang Quan Zhang

  3. Jiangsu Key Laboratory for Supermolecular Medicinal Materials and Applications, Nanjing Normal University, 210046, Nanjing, People’s Republic of China

    Shuang Quan Zhang

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  1. Jian Feng Li
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  2. Jie Zhang
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  3. Zhen Zhang
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  4. Hong Wei Ma
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Correspondence to Shuang Quan Zhang.

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Li, J.F., Zhang, J., Zhang, Z. et al. Production of Bioactive Human Beta-Defensin-4 in Escherichia coli Using SUMO Fusion Partner. Protein J 29, 314–319 (2010). https://doi.org/10.1007/s10930-010-9254-4

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  • DOI: https://doi.org/10.1007/s10930-010-9254-4

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