2. Academic Validation
  3. Soluble expression and purification of human β-defensin DEFB136 in Escherichia coli and identification of its bioactivity

Soluble expression and purification of human β-defensin DEFB136 in Escherichia coli and identification of its bioactivity

  • Protein Expr Purif. 2021 Dec;188:105968. doi: 10.1016/j.pep.2021.105968.
Haiyan Liu 1,  Hua Diao 2,  Jing Hou 3,  Heguo Yu 2,  Huiping Wen 1
Affiliations

Affiliations

  • 1 Department of Biology, College of Ecology, Lishui University, Lishui City, 323000, China.
  • 2 NPFPC Key Laboratory of Contraceptives and Devices, Shanghai Institute of Planned Parenthood Research, Shanghai, 200032, China.
  • 3 Department of Biology, College of Ecology, Lishui University, Lishui City, 323000, China. Electronic address: [email protected].
PMID: 34481960 DOI: 10.1016/j.pep.2021.105968
Abstract

Human β-defensins are an important family of innate host defense Peptides with pleiotropic activities. Human β-defensin 36 (DEFB136) is a novel member of the β-defensin family which have not been characterized so far. In the present research, the DEFB136 peptide was expressed successfully and purified using the IMPACT-TWIN 1 expression system. The purified DEFB136 peptide was identified by MALDI-TOF mass spectrometry and circular dichroism spectroscopy. While the recombinant DEFB136 peptide exhibited a broad spectrum of antimicrobial activity against E. coli, Staphylococcus aureus and Candida albicans strains, but had low cytotoxicity to human erythrocytes. In addition, the result of the octet assay showed that the DEFB136 had a high lipopolysaccharide (LPS)-binding affinity, suggesting the DEFB136 may be involved in immunoregulation through its LPS neutralization. These results may help lay the groundwork to understand better the complex interaction between innate host defense and the diversity of the defensin family.

Keywords

Antimicrobial activity; DEFB136; Human beta defensin; LPS; Soluble expression.

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