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Production of Bioactive Human β-defensin-3 in Escherichia coli by Soluble Fusion Expression

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Abstract

A codon optimized mature human β-defensin-3 gene (smHBD3) was synthesized and fused with TrxA to construct pET32-smHBD3 vector, which was transformed into E. coli BL21(DE3) and cultured in MBL medium. The volumetric productivity of fusion protein reached 0.99 g fusion protein l−1, i.e. 0.21 g mature HBD3 l−1. Ninety-six percentage of the fusion protein was in a soluble form and constituted about 45% of the total soluble protein. After cell disruption, the soluble fusion protein was separated by affinity chromatography and cleaved by enterokinase, and then the mature HBD3 was purified by cationic ion exchange chromatography. The overall recovery ratio of HBD3 was 43%. The purified mature HBD3 demonstrated antimicrobial activity against E. coli.

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Authors and Affiliations

  1. Institute of Bioengineering, Department of Chemical and Biological Engineering, Zhejiang University, 310027, Hangzhou, China

    Lei Huang, Zhixia Zhong, Li Peng, Haiqin Chen, Zhinan Xu & Peilin Cen

  2. School of Bioscience and Bioengineering, Southern China University of Technology, 510640, Guangzhou, China

    Jufang Wang

Authors
  1. Lei Huang
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  2. Jufang Wang
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  3. Zhixia Zhong
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  4. Li Peng
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  5. Haiqin Chen
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  6. Zhinan Xu
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  7. Peilin Cen
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Corresponding author

Correspondence to Zhinan Xu.

Additional information

Revisions requested 13 December 2005; Revisions received 24 January 2006

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Huang, L., Wang, J., Zhong, Z. et al. Production of Bioactive Human β-defensin-3 in Escherichia coli by Soluble Fusion Expression. Biotechnol Lett 28, 627–632 (2006). https://doi.org/10.1007/s10529-006-0024-5

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  • DOI: https://doi.org/10.1007/s10529-006-0024-5

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