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Research article

Biotechnology and Bioengineering (BB)  |  Enzyme, Protein Engineering, and Metabolic Engineering

Design, Characterization, and Antimicrobial Activity of a Novel Antimicrobial Peptide Derived from Bovine Lactophoricin

Ji-Sun Kim 1, Ji-Ho Joeng 1 and Yongae Kim 1*

Department of Chemistry, Hankuk University of Foreign Studies, Yongin 17035, Republic of Korea

Received: September 6, 2016; Accepted: January 19, 2017

J. Microbiol. Biotechnol. 2017; 27(4): 759-767

Published April 28, 2017 https://doi.org/10.4014/jmb.1609.09004

Copyright © The Korean Society for Microbiology and Biotechnology.

Abstract

Lactophoricin (LPcin), which is a part of proteose peptone isolated from bovine milk, is a cationic amphipathic α-helical antimicrobial peptide. Its truncated variants and mutated analogs were designed and their antimicrobial activities were evaluated by using various assays, like broth dilution methods and disk diffusion methods as well as hemolysis assay. Three analogs, LPcin-C8 (LPcin-YK1), LPcin-T2&6W (LPcin-YK2), and LPcin-T2&6W-C8 (LPcin-YK3), which showed better antibiotic activities than LPcin, were selected. Their secondary structures were also characterized by using CD spectropolarimetry. These three analogs of LPcin could be used as an alternative source of powerful antibacterial agents.

Keywords

Bovine lactophoricin, antimicrobial peptide, cationic amphipathic peptides, engineered analogs, antimicrobial activity

References

  1. Hancock REW. 2001. Cationic peptides: effectors in innate immunity and novel antimicrobials. Lancet Infect. Dis. 1: 156164.
    CrossRef
  2. Joerger RD. 2003. Alternatives to antibiotics: bacteriocins, antimicrobial peptides and bacteriophages. Poult. Sci. 82:640-647.
    Pubmed CrossRef
  3. Brad P. 2013. Washington Post. Available from https://www.washingtonpost.com/news/wonk/wp/2013/12/14/thefda-is-cracking-down-on-antibiotics-at-farms-heres-what-youshould-know/. Accessed December 14, 2013.
  4. Hassan M, Kjos M, Nes IF, Diep DB, Lotfipour F. 2012. Natural antimicrobial peptides from bacteria:characteristics and potential applications to fight against antibiotic resistance. J. Appl. Microbiol. 113: 723-736.
    Pubmed CrossRef
  5. Brown KL, Hancock REW. 2006. Cationic host defense (antimicrobial) peptides. Curr. Opin. Immunol. 18: 24-30.
    Pubmed CrossRef
  6. Friedrich CL, Moyles D, Beveridge TJ, Hancock REW. 2000. Antibacterial action of structurally diverse cationic peptides on gram-positive bacteria. Antimicrob. Agents Chemother. 44:2086-2092.
    Pubmed PMC CrossRef
  7. Ganz T, Selsted ME, Szklarek D, Harwig SS, Daher K, Bainton DF, Lehrer RI. 1985. Defensins. Natural peptide antibiotics of human neutrophils. J. Clin. Invest. 76: 1427-1435.
    Pubmed PMC CrossRef
  8. Hancock REW, Diamond G. 2000. The role of cationic antimicrobial peptides in innate host defences. Trends Microbiol. 8: 402-410.
    CrossRef
  9. Filipovića N, Borrmannb H, Todorovićc T, Borna M, Spasojevićd V, Sladićc D, et al. 2009. Copper(II) complexes of N-heteroaromatic hydrazones: synthesis, X-ray structure, magnetic behavior, and antibacterial activity. Inorg. Chim. Acta 362: 1996-2000.
    CrossRef
  10. Hancock REW, Rozek A. 2002. Role of membranes in the activities of antimicrobial cationic peptides. FEMS Microbiol. Lett. 206: 143-149.
    Pubmed CrossRef
  11. Bulet P, Stöcklin R, Menin L. 2004. Anti-microbial peptides:from invertebrates to vertebrates. Immunol. Rev. 198: 169-184.
    Pubmed CrossRef
  12. Steiner H, Hultmark D, Engström A, Bennich H, Boman HG. 1981. Sequence and specificity of two antibacterial proteins involved in insect immunity. Nature 292: 246-248.
    Pubmed CrossRef
  13. Andreu D, Rivas L. 1998. Animal antimicrobial peptides: an overview. Biopolymers 47: 415-433.
    CrossRef
  14. Huang HW. 2000. Action of antimicrobial peptides: twostate model. Biochemistry 39: 8347-8352.
    Pubmed CrossRef
  15. Matsuzaki K, Sugishita K, Fujii N, Miyajima K. 1995. Molecular basis for membrane selectivity of an antimicrobial peptide, magainin 2. Biochemistry 34: 3423-3429.
    Pubmed CrossRef
  16. Papagianni M. 2003. Ribosomally synthesized peptides with antimicrobial properties: biosynthesis, structure, function, and applications. Biotechnol. Adv. 21: 465-499.
    CrossRef
  17. Zhang L, Rozek A, Hancock REW. 2001. Interaction of cationic peptides with model membranes. J. Biol. Chem. 276:35714-35722.
    Pubmed CrossRef
  18. Mohammad FV, Noorwala M, Ahmad VU, Sener B. 1995. Bidesmosidic triterpenoidal saponins from the roots of Symphytum officinale. Planta Med. 61: 94.
    Pubmed CrossRef
  19. Aley SB, Zimmerman M, Hetsko M, Selsted ME, Gillin FD. 1994. Killing of Giardia lamblia by cryptdins and cationic neutrophil peptides. Infect. Immun. 62: 5397-5403.
    Pubmed PMC
  20. Campagna S, Mathot AG, Fleury Y, Girardet JM, Gaillard JL. 2004. Antibacterial activity of lactophoricin, a synthetic 23-residues peptide derived from the sequence of bovine milk component-3 of proteose peptone. J. Dairy Sci. 87:1621-1626.
    CrossRef
  21. Gor’kov PL, Chekmenev EY, Li C, Cotten M, Buffy Jarrod J, Traaseth Nathaniel J, et al. 2007. Using low-E resonators to reduce RF heating in biological samples for static solid-state NMR up to 900 MHz. J. Magn. Reson. 185: 77-93.
    Pubmed CrossRef
  22. Park TJ, Kim JS, Choi SS, Kim Y. 2009. Cloning expression, isotope labeling, purification and characterization of bovine antimicrobial peptide, lactophoricin in Escherichia coli. Protein Expr. Purif. 65: 23-29.
    Pubmed CrossRef
  23. Park TJ, Kim JS, Ahn HC, Kim Y. 2011. Solution and solidstate NMR structural studies of antimicrobial peptides LPcin-I and LPcin-II. Biophys. J. 101: 1193-1201.
    Pubmed PMC CrossRef
  24. Bechinger B. 1997. Structure and functions of channel-forming peptides: magainins, cecropins, melittin and alamethicin. J. Membr. Biol. 156: 197-211.
    Pubmed CrossRef
  25. Hancock REW, Lehrer R. 1998. Cationic peptides: a new source of antibiotics. Trends Biotechnol. 16: 82-88.
    CrossRef
  26. Fjell CD, Hiss JA, Hancock REW, Schneider G. 2012. Designing antimicrobial peptides: form follows function. Nat. Rev. Drug Discov. 11: 37-51.
    CrossRef
  27. Tossi A, Tarantino C, Romeo D. 1997. Design of synthetic antimicrobial peptides based on sequence analogy and amphipathicity. Eur. J. Biochem. 250: 549-558.
    Pubmed CrossRef
  28. Wu G, Ding J, Li H, Li L, Zhao R, Shen Z, et al. 2008. Effects of cations and pH on antimicrobial activity of thanatin and s-thanatin against Escherichia coli ATCC25922 and B. subtilis ATCC 21332. Curr. Microbiol. 57: 552-557.
    Pubmed CrossRef
  29. Kim JS, Jeong JH, Kim KS, Kim Y. 2015. Optimized expression and characterization of antimicrobial peptides, LPcin analogs. Bull. Korean Chem. Soc. 36: 1148-1154.
    CrossRef
  30. Mishra B, Basu A, Chua RRY, Saravanan R, Tambyah PA, Ho B, et al. 2014. Site specific immobilization of a potent antimicrobial peptide onto silicone catheters: evaluation against urinary tract infection pathogens. J. Mater. Chem. B 2: 1706-1716.
    CrossRef
  31. Greenfield NJ. 2006. Using circular dichroism spectra to estimate protein secondary structure. Nat. Protoc. 1: 2876-2890.
    Pubmed PMC CrossRef
  32. Gallo RL, Murakami M, Ohtake T, Zaiou M. 2002. Biology and clinical relevance of naturally occurring antimicrobial peptides. J. Allergy Clin. Immunol. 110: 823-831.
    Pubmed CrossRef
  33. Hancock REW, Sahl HG. 2006. Antimicrobial and hostdefense peptides as new anti-infective therapeutic strategies. Nat. Biotechnol. 24: 1551-1557.
    Pubmed CrossRef
  34. Tossi A, Sandri L, Giangaspero A. 2000. Amphipathic, alpha-helical antimicrobial peptides. Biopolymers 55: 4-30.
    CrossRef
  35. Jeong JH, Kim JS, Choi SS, Kim Y. 2016. NMR structural studies of antimicrobial peptides: LPcin analogs. Biophys. J. 110: 423-430.
    Pubmed PMC CrossRef

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Article

Research article

J. Microbiol. Biotechnol. 2017; 27(4): 759-767

Published online April 28, 2017 https://doi.org/10.4014/jmb.1609.09004

Copyright © The Korean Society for Microbiology and Biotechnology.

Design, Characterization, and Antimicrobial Activity of a Novel Antimicrobial Peptide Derived from Bovine Lactophoricin

Ji-Sun Kim 1, Ji-Ho Joeng 1 and Yongae Kim 1*

Department of Chemistry, Hankuk University of Foreign Studies, Yongin 17035, Republic of Korea

Received: September 6, 2016; Accepted: January 19, 2017

Abstract

Lactophoricin (LPcin), which is a part of proteose peptone isolated from bovine milk, is a
cationic amphipathic α-helical antimicrobial peptide. Its truncated variants and mutated
analogs were designed and their antimicrobial activities were evaluated by using various
assays, like broth dilution methods and disk diffusion methods as well as hemolysis assay.
Three analogs, LPcin-C8 (LPcin-YK1), LPcin-T2&6W (LPcin-YK2), and LPcin-T2&6W-C8
(LPcin-YK3), which showed better antibiotic activities than LPcin, were selected. Their
secondary structures were also characterized by using CD spectropolarimetry. These three
analogs of LPcin could be used as an alternative source of powerful antibacterial agents.

Keywords: Bovine lactophoricin, antimicrobial peptide, cationic amphipathic peptides, engineered analogs, antimicrobial activity

References

  1. Hancock REW. 2001. Cationic peptides: effectors in innate immunity and novel antimicrobials. Lancet Infect. Dis. 1: 156164.
    CrossRef
  2. Joerger RD. 2003. Alternatives to antibiotics: bacteriocins, antimicrobial peptides and bacteriophages. Poult. Sci. 82:640-647.
    Pubmed CrossRef
  3. Brad P. 2013. Washington Post. Available from https://www.washingtonpost.com/news/wonk/wp/2013/12/14/thefda-is-cracking-down-on-antibiotics-at-farms-heres-what-youshould-know/. Accessed December 14, 2013.
  4. Hassan M, Kjos M, Nes IF, Diep DB, Lotfipour F. 2012. Natural antimicrobial peptides from bacteria:characteristics and potential applications to fight against antibiotic resistance. J. Appl. Microbiol. 113: 723-736.
    Pubmed CrossRef
  5. Brown KL, Hancock REW. 2006. Cationic host defense (antimicrobial) peptides. Curr. Opin. Immunol. 18: 24-30.
    Pubmed CrossRef
  6. Friedrich CL, Moyles D, Beveridge TJ, Hancock REW. 2000. Antibacterial action of structurally diverse cationic peptides on gram-positive bacteria. Antimicrob. Agents Chemother. 44:2086-2092.
    Pubmed KoreaMed CrossRef
  7. Ganz T, Selsted ME, Szklarek D, Harwig SS, Daher K, Bainton DF, Lehrer RI. 1985. Defensins. Natural peptide antibiotics of human neutrophils. J. Clin. Invest. 76: 1427-1435.
    Pubmed KoreaMed CrossRef
  8. Hancock REW, Diamond G. 2000. The role of cationic antimicrobial peptides in innate host defences. Trends Microbiol. 8: 402-410.
    CrossRef
  9. Filipovića N, Borrmannb H, Todorovićc T, Borna M, Spasojevićd V, Sladićc D, et al. 2009. Copper(II) complexes of N-heteroaromatic hydrazones: synthesis, X-ray structure, magnetic behavior, and antibacterial activity. Inorg. Chim. Acta 362: 1996-2000.
    CrossRef
  10. Hancock REW, Rozek A. 2002. Role of membranes in the activities of antimicrobial cationic peptides. FEMS Microbiol. Lett. 206: 143-149.
    Pubmed CrossRef
  11. Bulet P, Stöcklin R, Menin L. 2004. Anti-microbial peptides:from invertebrates to vertebrates. Immunol. Rev. 198: 169-184.
    Pubmed CrossRef
  12. Steiner H, Hultmark D, Engström A, Bennich H, Boman HG. 1981. Sequence and specificity of two antibacterial proteins involved in insect immunity. Nature 292: 246-248.
    Pubmed CrossRef
  13. Andreu D, Rivas L. 1998. Animal antimicrobial peptides: an overview. Biopolymers 47: 415-433.
    CrossRef
  14. Huang HW. 2000. Action of antimicrobial peptides: twostate model. Biochemistry 39: 8347-8352.
    Pubmed CrossRef
  15. Matsuzaki K, Sugishita K, Fujii N, Miyajima K. 1995. Molecular basis for membrane selectivity of an antimicrobial peptide, magainin 2. Biochemistry 34: 3423-3429.
    Pubmed CrossRef
  16. Papagianni M. 2003. Ribosomally synthesized peptides with antimicrobial properties: biosynthesis, structure, function, and applications. Biotechnol. Adv. 21: 465-499.
    CrossRef
  17. Zhang L, Rozek A, Hancock REW. 2001. Interaction of cationic peptides with model membranes. J. Biol. Chem. 276:35714-35722.
    Pubmed CrossRef
  18. Mohammad FV, Noorwala M, Ahmad VU, Sener B. 1995. Bidesmosidic triterpenoidal saponins from the roots of Symphytum officinale. Planta Med. 61: 94.
    Pubmed CrossRef
  19. Aley SB, Zimmerman M, Hetsko M, Selsted ME, Gillin FD. 1994. Killing of Giardia lamblia by cryptdins and cationic neutrophil peptides. Infect. Immun. 62: 5397-5403.
    Pubmed KoreaMed
  20. Campagna S, Mathot AG, Fleury Y, Girardet JM, Gaillard JL. 2004. Antibacterial activity of lactophoricin, a synthetic 23-residues peptide derived from the sequence of bovine milk component-3 of proteose peptone. J. Dairy Sci. 87:1621-1626.
    CrossRef
  21. Gor’kov PL, Chekmenev EY, Li C, Cotten M, Buffy Jarrod J, Traaseth Nathaniel J, et al. 2007. Using low-E resonators to reduce RF heating in biological samples for static solid-state NMR up to 900 MHz. J. Magn. Reson. 185: 77-93.
    Pubmed CrossRef
  22. Park TJ, Kim JS, Choi SS, Kim Y. 2009. Cloning expression, isotope labeling, purification and characterization of bovine antimicrobial peptide, lactophoricin in Escherichia coli. Protein Expr. Purif. 65: 23-29.
    Pubmed CrossRef
  23. Park TJ, Kim JS, Ahn HC, Kim Y. 2011. Solution and solidstate NMR structural studies of antimicrobial peptides LPcin-I and LPcin-II. Biophys. J. 101: 1193-1201.
    Pubmed KoreaMed CrossRef
  24. Bechinger B. 1997. Structure and functions of channel-forming peptides: magainins, cecropins, melittin and alamethicin. J. Membr. Biol. 156: 197-211.
    Pubmed CrossRef
  25. Hancock REW, Lehrer R. 1998. Cationic peptides: a new source of antibiotics. Trends Biotechnol. 16: 82-88.
    CrossRef
  26. Fjell CD, Hiss JA, Hancock REW, Schneider G. 2012. Designing antimicrobial peptides: form follows function. Nat. Rev. Drug Discov. 11: 37-51.
    CrossRef
  27. Tossi A, Tarantino C, Romeo D. 1997. Design of synthetic antimicrobial peptides based on sequence analogy and amphipathicity. Eur. J. Biochem. 250: 549-558.
    Pubmed CrossRef
  28. Wu G, Ding J, Li H, Li L, Zhao R, Shen Z, et al. 2008. Effects of cations and pH on antimicrobial activity of thanatin and s-thanatin against Escherichia coli ATCC25922 and B. subtilis ATCC 21332. Curr. Microbiol. 57: 552-557.
    Pubmed CrossRef
  29. Kim JS, Jeong JH, Kim KS, Kim Y. 2015. Optimized expression and characterization of antimicrobial peptides, LPcin analogs. Bull. Korean Chem. Soc. 36: 1148-1154.
    CrossRef
  30. Mishra B, Basu A, Chua RRY, Saravanan R, Tambyah PA, Ho B, et al. 2014. Site specific immobilization of a potent antimicrobial peptide onto silicone catheters: evaluation against urinary tract infection pathogens. J. Mater. Chem. B 2: 1706-1716.
    CrossRef
  31. Greenfield NJ. 2006. Using circular dichroism spectra to estimate protein secondary structure. Nat. Protoc. 1: 2876-2890.
    Pubmed KoreaMed CrossRef
  32. Gallo RL, Murakami M, Ohtake T, Zaiou M. 2002. Biology and clinical relevance of naturally occurring antimicrobial peptides. J. Allergy Clin. Immunol. 110: 823-831.
    Pubmed CrossRef
  33. Hancock REW, Sahl HG. 2006. Antimicrobial and hostdefense peptides as new anti-infective therapeutic strategies. Nat. Biotechnol. 24: 1551-1557.
    Pubmed CrossRef
  34. Tossi A, Sandri L, Giangaspero A. 2000. Amphipathic, alpha-helical antimicrobial peptides. Biopolymers 55: 4-30.
    CrossRef
  35. Jeong JH, Kim JS, Choi SS, Kim Y. 2016. NMR structural studies of antimicrobial peptides: LPcin analogs. Biophys. J. 110: 423-430.
    Pubmed KoreaMed CrossRef