Purification of Antibacterial Peptide from the Skin of the Catfish Silurus asotus

메기의 껍질로부터 항균성 펩타이드의 정제

  • Received : 2015.12.02
  • Accepted : 2016.02.06
  • Published : 2016.03.30


An antibacterial peptide from skin extract of the catfish Silurus asotus was purified and characterized. The acidified skin extract was put through a Sep-Pak C18 solid phase extraction cartridge using a stepwise gradient and divided into flow-through (F.T.), 10% methanol-elute (RM10), 60% methanolelute (RM60), and 100% methanol-elute (RM100) fractions. RM10, RM60, and RM 100 showed antimicrobial activity against Escherichia coli D31. On the other hand, the F.T. fraction did not show antimicrobial activity. Among the various fractions, RM 60 had the highest activity. RM 60 was partially purified on a cation exchange column (CM52) by a stepwise gradient. The ammonium acetate (pH 5.15) 0.02 M – 0.8 M fraction showed antimicrobial activity. Then an antimicrobial peptide was purified using a 0.6M fraction with strong antibacterial activity through a series of five C18 reversed-phase HPLC columns. For the characterization of the purified peptide, the molecular weight and amino acid sequence were analyzed by MALDI-TOF MS and Edman degradation. The molecular weight of this peptide was about 4182.1 [M+H]+. The amino acid sequence of this peptide was partially determined as follows: PALXXKARREAKVKF. These findings suggest that this peptide plays a significant role in the innate defense system of catfish skin.


Antibacterial peptide;catfish;isolation;skin


  1. Acosta, J., Montero, V., Carpio, Y., Velázquez, J., Garay, H. E., Reyes, O., Cabrales, A., Masforrol, Y., Morales, A. and Estrada, M. P. 2013. Cloning and functional characterization of three novel antimicrobial peptides from tilapia (Oreochromis niloticus). Aquaculture 372-375, 9-18.
  2. Bartlett, T. C., Cuthbertson, B. J., Shepard, E. F., Chapman, R. W., Gross, P. S. and Warr, G. W. 2002. Crustins, homologues of an 11.5-kDa antibacterial peptide, from two species of penaeid shrimp, Litopenaeus vannamei and Litopenaeus setiferus. Mar. Biotechnol. 4, 278-293.
  3. Bulet, P., Stöcklin, R. and Menin, L. 2004. Anti-microbial peptides: from invertebrates to vertebrates. Immunol. Rev. 198, 169-184.
  4. Cole, A. M., Weis, P. and Diamond, G. 1997. Isolation and characterization of pleurocidin, an antimicrobial peptide in the skin secretions of winter flounder. J. Biol. Chem. 272, 12008-12013.
  5. Ellis, A. E. 2001. Innate host defense mechanisms of fish against viruses and bacteria. Dev. Comp. Immunol. 25, 827- 839.
  6. Ferraro, V., Cruz, I. B., Jorge, R. F., Malcata, F. X., Pintado, M. E. and Castro, P. M. L. 2010. Valorization of natural extracts from marine source focused on marine byproducts: a review. Food Res. Int. 43, 2221-2233.
  7. Go, H.-J., Bae, Y. J. and Park, N. G. 2014. Purification of two novel antimicrobial peptides from pyloric caeca of the starfish Asterina pectinifera. J. Life Sci. 24, 860-864.
  8. House, R. V. and Hastings, K. L. 2004. Opinion in immunotoxicology: multidimensional immunemodula-tion. J. Immunotoxicol. 1, 123-129.
  9. Izadpanah, A. and Gallo, R. L. 2005. Antimicrobial peptides. J. Am. Acad. Dermatol. 52, 381-390.
  10. Jenssen, H., Hamill, P. and Hancock, R. E. 2006. Peptide antimicrobial agents. Clin. Microbiol. Rev. 19, 491-511.
  11. Klomklao, S., Kishimura, H., Nonami, Y. and Benjakul, S. 2009. Biochemical properties of two isoforms of trypsin purified from the Intestine of skipjack tuna (Katsuwonus pelamis). Food Chem. 115, 155-162.
  12. Oren, Z. and Shai, Y. 1996. A class of highly potent antibacterial peptides related from pardaxin, a pore-forming peptide from the Moses sole fish Pardachirus marmoratus. Eur. J. Biochem. 237, 304-310.
  13. Ovchinnikova, T. V., Aleshina, G. M., Balandin, S. V., Krasnosdembskaya, A. D., Markelov, M. L., Frolova, E. I., Leonova, Y. F., Tagaev, A. A., Krasnodembsky, E. G. and Kokryakov, V. N. 2004. Purification and primary structure of two isoforms of arenicin, a novel antimicrobial peptide from marine polychaeta Arenicola marina. FEBS Lett. 577, 209-214.
  14. Park, C. B., Lee, J. H., Park, I. Y., Kim, M. S. and Kim, S. C. 1997. A novel antimicrobial peptide from the loach, Misgurnus anguillicaudatus. FEBS Lett. 411, 173-178.
  15. Park, I. Y., Park, C. B., Kim, M. S. and Kim, S. C. 1998. Parasin I, an antimicrobial peptide derived from histone H2A in the catfish, Parasilurus asotus. FEBS Lett. 437, 258-262.
  16. Pasupuleti, M., Schmidtchen, A. and Malmsten, M. 2012. Antimicrobial peptides: key components of the innate immune system. Crit. Rev. Biotechnol. 32, 143-171.
  17. Rakers, S., Niklasson l., Steinhagen D. Kruse, C., Schauber J., Sundell, K. and Paus, R. 2013. Antimicrobial peptides (AMPs) from fish epidermis: perspectives for investigative dermatology. J. Invest. Dermatol. 133, 1140-1149.
  18. Seo, J. K., Crawford, J. M., Stone, K. L. and Noga, E. J. 2005. Purification of a novel arthropod defensin from the American oyster, Crassostrea virginica. Biochem. Biophys. Res. Commun. 338, 1998-2004.
  19. Seo, J. K., Lee, M. J., Go, H. J., Park, T. H. and Park, N. G. 2012. Purification and characterization of YFGAP, a GAPDH-related novel antimicrobial peptide, from the skin of yellowfin tuna, Thunnus albacares. Fish. Shellfish Immunol. 33, 743-752.
  20. Shike, H., Lauth, X., Westerman, M. E., Ostland, V. E., Carlberg, J. M., Van Olst, J. C., Shimizu, C., Bulet, P. and Burns, J. C. 2002. Bass hepcidin is a novel antimicrobial peptide induced by bacterial challenge. Eur. J. Biochem. 269, 2232-2237.
  21. Silphaduang, U. and Noga, E. J. 2001. Peptide antibiotics in mast cells of fish. Nature 414, 268-269.
  22. Stark, M., Liu, L. P. and Deber, C. M. 2002. Cationic Hydrophobic Peptides with Antimicrobial Activity. Antimicrob. Agents Chemother. 46, 3585-3590.
  23. Subramanian, S., Ross, N. W. and MacKinnon, S. L. 2009. Myxinidin, a novel antimicrobial peptide from the epidermal mucus of hagfish, Myxine glutinosa L. Mar. Biotechnol. 11, 748-757.
  24. Ullal, A. J., Litaker, R. W. and Noga, E. J. 2008. Antimicrobial peptides related from hemoglobin are expressed in epithelium of channel catfish (Ictalurus punctatus, Rafinesque). Dev. Comp. Immunol. 32, 1301-1312.
  25. van’t Hof, W., Veerman, E. C., Helmerhorst, E. J. and Amerongen, A. V. 2001. Antimicrobial peptides: properties and applicability. Biol. Chem. 382, 597-619.