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Article

Secretion and activity of antimicrobial peptide cecropin D expressed in Pichia pastoris

  • Authors:
    • Chunhe Guo
    • Yumao Huang
    • Hongyu Zheng
    • Liyun Tang
    • Jun  He
    • Linsheng Xiang
    • Dehui Liu
    • Houquan Jiang
  • View Affiliations / Copyright

    Affiliations: College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong 510642, P.R. China
  • Pages: 1063-1068
    |
    Published online on: September 24, 2012
       https://doi.org/10.3892/etm.2012.719
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Abstract

To express the antimicrobial peptide cecropin D in Pichia pastoris and determine the activity of the expressed product, four oligonucleotide fragments were synthesized in accordance with the available cecropin D sequences and a codon bias suitable for Pichia pastoris. Sequence fragments were phosphorylated, annealed, linked and cloned into the expression vector pGAPZαA and the yeast α‑mating factor signal peptide was used as the signal sequence. The P. pastoris SMD1168 cells were transformed by electroporation using the constructed recombinant plasmid pGAPZαA‑cecropin D. We were able to demonstrate by PCR that the cecropin D sequence had integrated into the P. pastoris genome. The expressed and secreted product was identified using Tricine‑SDS‑PAGE. Antibacterial activity was demonstrated using an agarose diffusion test and turbidimetry. The molecular mass of the recombinant cecropin D was estimated to be 3,900 Da. The recombinant cecropin D exhibited antibacterial activity for both Gram‑positive and Gram‑negative bacteria, suggesting that cecropin D was successfully expressed in P. pastoris. This approach holds great promise for antibacterial drug development.
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Copy and paste a formatted citation
Spandidos Publications style
Guo C, Huang Y, Zheng H, Tang L, He J, Xiang L, Liu D and Jiang H: Secretion and activity of antimicrobial peptide cecropin D expressed in Pichia pastoris. Exp Ther Med 4: 1063-1068, 2012.
APA
Guo, C., Huang, Y., Zheng, H., Tang, L., He, J., Xiang, L. ... Jiang, H. (2012). Secretion and activity of antimicrobial peptide cecropin D expressed in Pichia pastoris. Experimental and Therapeutic Medicine, 4, 1063-1068. https://doi.org/10.3892/etm.2012.719
MLA
Guo, C., Huang, Y., Zheng, H., Tang, L., He, J., Xiang, L., Liu, D., Jiang, H."Secretion and activity of antimicrobial peptide cecropin D expressed in Pichia pastoris". Experimental and Therapeutic Medicine 4.6 (2012): 1063-1068.
Chicago
Guo, C., Huang, Y., Zheng, H., Tang, L., He, J., Xiang, L., Liu, D., Jiang, H."Secretion and activity of antimicrobial peptide cecropin D expressed in Pichia pastoris". Experimental and Therapeutic Medicine 4, no. 6 (2012): 1063-1068. https://doi.org/10.3892/etm.2012.719
Copy and paste a formatted citation
x
Spandidos Publications style
Guo C, Huang Y, Zheng H, Tang L, He J, Xiang L, Liu D and Jiang H: Secretion and activity of antimicrobial peptide cecropin D expressed in Pichia pastoris. Exp Ther Med 4: 1063-1068, 2012.
APA
Guo, C., Huang, Y., Zheng, H., Tang, L., He, J., Xiang, L. ... Jiang, H. (2012). Secretion and activity of antimicrobial peptide cecropin D expressed in Pichia pastoris. Experimental and Therapeutic Medicine, 4, 1063-1068. https://doi.org/10.3892/etm.2012.719
MLA
Guo, C., Huang, Y., Zheng, H., Tang, L., He, J., Xiang, L., Liu, D., Jiang, H."Secretion and activity of antimicrobial peptide cecropin D expressed in Pichia pastoris". Experimental and Therapeutic Medicine 4.6 (2012): 1063-1068.
Chicago
Guo, C., Huang, Y., Zheng, H., Tang, L., He, J., Xiang, L., Liu, D., Jiang, H."Secretion and activity of antimicrobial peptide cecropin D expressed in Pichia pastoris". Experimental and Therapeutic Medicine 4, no. 6 (2012): 1063-1068. https://doi.org/10.3892/etm.2012.719
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