International Journal of Industrial Entomology and Biomaterials

Korean Society of Sericultural Science (한국잠사학회)
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Antibacterial effects of two cecropin type peptides isolated from the silkworm against Salmonella species

  • Kim, Seong Ryul (Department of Agricultural Biology, National Institute of Agricultural Science, RDA,) ;
  • Park, Jong Woo (Department of Agricultural Biology, National Institute of Agricultural Science, RDA,) ;
  • Kim, Seong-Wan (Department of Agricultural Biology, National Institute of Agricultural Science, RDA,) ;
  • Kim, Su Bae (Department of Agricultural Biology, National Institute of Agricultural Science, RDA,) ;
  • Jo, You-Young (Department of Agricultural Biology, National Institute of Agricultural Science, RDA,) ;
  • Kim, Kee Young (Department of Agricultural Biology, National Institute of Agricultural Science, RDA,) ;
  • Choi, Kwang-Ho (Department of Agricultural Biology, National Institute of Agricultural Science, RDA,) ;
  • Ji, Sang Deok (Department of Agricultural Biology, National Institute of Agricultural Science, RDA,) ;
  • Kim, Jong gil (Department of Agricultural Biology, National Institute of Agricultural Science, RDA,) ;
  • Kweon, HaeYong (Department of Agricultural Biology, National Institute of Agricultural Science, RDA,)
  • Received : 2018.11.15
  • Accepted : 2018.12.04
  • Published : 2018.12.31
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Abstract

In insect defense system, antimicrobial peptides (AMPs) are one of important biological molecules to survive in a variety of environments. Insect can synthesize AMPs to protect against invading pathogens in humoral immune response. Taking more advantage of biological antimicrobial molecules, we report antibacterial activity of two cecropin type peptides, cecropin and moricin, isolated from the silkworm against four salmonella species. In this work, we purified antimicrobial candidate peptides (AMCP) from the extracts of immune challenged silkworm larval hemolymph by two-step chromatographic purification procedure, cation exchange and gel permeation chromatography. The molecular weights of purified peptides were estimated to be about 4 ~ 5 kDa by Tricin SDS-PAGE analysis, and identified as silkworm cecropin and moricin by NCBI BLAST homology search with their N-terminal amino acid sequences. As antibacterial activity assay, the purified peptides showed stronger antibacterial activity against Salmonella pathogens with an MIC value of $1{\sim}4{\mu}g/mL$. Therefore two cecropin type peptides purified from the silkworm will be valuable potential materials for development of new natural antibiotics.

Keywords

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Fig. 1. Fractionation of hemolymph extracts from the immunized larvae by cation exchange chromatography. (A) Profile of cation exchange chromatography using HiTrap SP HP column with linear gradient of 0 - 80% 1.5M NaCl as the first step purification. The peak containing antibacterial activity is indicated with an arrow. (B) Antibacterial activity of fractions eluted form the cation exchange column by agar well diffusion assay (C) Tricine SDS-PAGE analysis of eluted fractions containing antibacterial activity.

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Fig. 2. Final purification of antimicrobial peptides by gel permeation chromatography. (A) Profile of superdex peptide gel permeation chromatography as the final purification. The peaks containing strongest antibacterial activity are indicated with arrows. (B) Agar well diffusion assay of fractions 19 ~30 collected form the superdex peptide column. (C) Tricine SDS-PAGE analysis of fractions 18 ~ 28 containing antibacterial activity.

Table 1. N-terminal amino acid sequences of purifed peptides

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Table 2. Antimicrobial activity of purifed cecropin and moricin

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References

  1. Bulet P, Stocklin R (2005) Insect antimicrobial peptides: structures, properties and gene regulation. Protein Pept Lett 12, 3-11. https://doi.org/10.2174/0929866053406011
  2. Dai H, Rayaprolu S, Gong Y, Huang R, Prakash O, Jiang H (2008) Solution structure, antibacterial activity, and expression profile of Manduca sexta moricin. J Pep Sci 14, 855-863. https://doi.org/10.1002/psc.1016
  3. Hara S, Yamakawa M (1995) Moricin, a novel type of antibacterial peptide isolated from the silkworm, Bombyx mori. Biochem Biophys Res Commun 220, 664-669.
  4. Hoffman JA, Kafatos FC, Janeway CA, Ezekowitz RA (1999) Phylogenetic perspectives in innate immunity. Science 284, 1313-1318. https://doi.org/10.1126/science.284.5418.1313
  5. Jenssen H, Hamill P, Hancock RE (2006) Peptide antimicrobial agents, Clin Microbiol Rev 19, 491-511. https://doi.org/10.1128/CMR.00056-05
  6. Kim JK, Lee E, Shin S, Jeong KW, Lee JY, Bae SY, et al. (2011) Structure and function of papiliocin with antimicrobial and antiinflammatory activities isolated from the swallowtail butterfly, Papilio xuthus. J Biol Chem 286, 41296-41311. https://doi.org/10.1074/jbc.M111.269225
  7. Kim SR, Choi KH, Kim SW, Hwang JS, Goo TW, Kim I (2015) Molecular cloning of a novel cecropin-like peptide gene from the swallowtail butterfly, papilo Xuthus. Int J Indust Entomol 31(1), 1-6. https://doi.org/10.7852/ijie.2015.31.1.1
  8. Kim SR, Choi KH, Kim SW, Park SW (2016) Comparison of gloverin gene expression patterns between domesticated and wild silkworms. Int J Indust Entomol 33(2), 1-8. https://doi.org/10.7852/ijie.2016.33.1.1
  9. Kim SR, Hong MY, Park SW, Choi KH, Yun EY, Goo TW, et al. (2010) Characterization and cDNA cloning of cecropin-like antimicrobial peptide, Papiliocin from the swallowtail butterfly, Papilio xuthus. Mol Cells 29, 419-423. https://doi.org/10.1007/s10059-010-0050-y
  10. Steiner H, Hultmark D, Engstrom A, Bennich H, Boman HG (1981) Sequence and specificity of two antibacterial proteins involved in insect immunity. Nature 292, 246-248. https://doi.org/10.1038/292246a0
  11. Suttmann H, Retz M, Paulsen F, Harder J, Zwergel U, Kamradt J, et al. (2008) Antimicrobial peptides of the Cecropin-family show potent antitumor activity against bladder cancer cells. BMC Urol 8,5. https://doi.org/10.1186/1471-2490-8-5
  12. Wachinger M, Kleinschmidt A, Winder D, von Pechmann N, Ludvigsen A, Neumann M, et al. (1998) Antimicrobial peptides melittin and cecropin inhibit replication of human immunodeficiency virus 1 by suppressing viral gene expression. J Gen Virol 79(Pt 4), 731-740. https://doi.org/10.1099/0022-1317-79-4-731
  13. Yi HY, Chowdhury M, Huang YD, Yu XQ (2014) Insect antimicrobial peptides and their applications. Appl Microbiol Biotechnol 98(13), 5807-5822. https://doi.org/10.1007/s00253-014-5792-6

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