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http://dx.doi.org/10.5713/ajas.18.0571

Identification of duck liver-expressed antimicrobial peptide 2 and characterization of its bactericidal activity  

Hong, Yeojin (Department of Animal Science and Technology, Chung-Ang University)
Truong, Anh Duc (Department of Animal Science and Technology, Chung-Ang University)
Lee, Janggeun (Department of Animal Science and Technology, Chung-Ang University)
Lee, Kyungbaek (Department of Animal Science and Technology, Chung-Ang University)
Kim, Geun-Bae (Department of Animal Science and Technology, Chung-Ang University)
Heo, Kang-Nyeong (Poultry Research Institute, National Institute of Animal Science, RDA)
Lillehoj, Hyun S. (Animal Biosciences and Biotechnology Laboratory, Agricultural Research Services, United States Department of Agriculture)
Hong, Yeong Ho (Department of Animal Science and Technology, Chung-Ang University)
Publication Information
Asian-Australasian Journal of Animal Sciences / v.32, no.7, 2019 , pp. 1052-1061 More about this Journal
Abstract
Objective: This study was conducted to identify duck liver-expressed antimicrobial peptide 2 (LEAP-2) and demonstrate its antimicrobial activity against various pathogens. Methods: Tissue samples were collected from 6 to 8-week-old Pekin ducks (Anas platyrhynchos domesticus), total RNA was extracted, and cDNA was synthesized. To confirm the duck LEAP-2 transcript expression levels, quantitative real-time polymerase chain reaction was conducted. Two kinds of peptides (a linear peptide and a disulfide-type peptide) were synthesized to compare the antimicrobial activity. Then, antimicrobial activity assay and fluorescence microscopic analysis were conducted to demonstrate duck LEAP-2 bactericidal activity. Results: The duck LEAP-2 peptide sequence showed high identity with those of other avian species (>85%), as well as more than 55% of identity with mammalian sequences. LEAP-2 mRNA was highly expressed in the liver with duodenum next, and then followed by lung, spleen, bursa and jejunum and was the lowest in the muscle. Both of LEAP-2 peptides efficiently killed bacteria, although the disulfide-type LEAP-2 showed more powerful bactericidal activity. Also, gram-positive bacteria was more susceptible to duck LEAP-2 than gram-negative bacteria. Using microscopy, we confirmed that LEAP-2 peptides could kill bacteria by disrupting the bacterial cell envelope. Conclusion: Duck LEAP-2 showed its antimicrobial activity against both gram-positive and gram-negative bacteria. Disulfide bonds were important for the powerful killing effect by disrupting the bacterial cell envelope. Therefore, duck LEAP-2 can be used for effective antibiotics alternatives.
Keywords
Antimicrobial Peptides; Liver-expressed Antimicrobial Peptide 2 (LEAP-2); Duck; Disulfide Bond; Pathogens;
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