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Expression of the zebrafish β-defensin 3 mature peptide in Pichia pastoris and its purification and antibacterial activity

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Abstract

Defensins are abundant in cells and tissues that are involved in host defense against microbial infections. zfDB3 (zebrafish β-defensin 3) is one of 3 copies of defensin β-like genes from zebrafish (Danio rerio). Here we focus on mzfDB3, which is the gene encoding for the zebrafish β-defensin 3 mature peptide. A codon-optimized mzfDB3 gene with a 6×His-tag at the 3′-end was inserted into the pPICZαA expression vector and transformed into Pichia pastoris X-33 cells. The recombinant zebrafish β-defensin 3 mature peptide (rmzfDB3) was induced with 1.0% methanol at 29°C for 72 h and purified by immobilized metal affinity chromatography. MALDI-TOF/TOF analysis confirmed the expected purified product (rmzfDB3, 5.9 kDa). Fermentation supernatant, which contained rmzfDB3, showed antibacterial activity against Grampositive (i.e., Listeria monocytogenes, Staphylococcus aureus, and Bacillus cereus) and Gram-negative (i.e., Escherchia coli BL21, Vibrio parahaemolyticus, Salmonella lignieres, and Pseudomonas aeruginosa) bacteria.

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Abbreviations

zfDB3:

zebrafish ß-defensin 3

mzfDB3:

zebrafish ß-defensin 3 mature peptide

rmzfDB3:

recombinant zebrafish ß-defensin 3 mature peptide

IMAC:

immobilized metal affinity chromatography

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Authors and Affiliations

  1. College of Food Science and Technology, Shanghai Ocean University, Shanghai, 201306, China

    Y. Tao, C. F. Song & W. Li

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  1. Y. Tao
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  2. C. F. Song
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  3. W. Li
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Correspondence to Y. Tao.

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Tao, Y., Song, C.F. & Li, W. Expression of the zebrafish β-defensin 3 mature peptide in Pichia pastoris and its purification and antibacterial activity. Appl Biochem Microbiol 53, 661–668 (2017). https://doi.org/10.1134/S0003683817060126

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  • DOI: https://doi.org/10.1134/S0003683817060126

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