Journal of Gastrointestinal Infections

Register      Login

VOLUME 2 , ISSUE 1 ( 2012 ) > List of Articles


Stability of antimicrobial activity of cryptdin-2 against selected pathogens under physiological conditions

Manesh Lahori, Praveen Rishi, Ram Prakash Tiwari

Keywords : antimicrobial activity, bacterial pathogens, cryptdin-2, physiological conditions, stability.

Citation Information : Lahori M, Rishi P, Tiwari RP. Stability of antimicrobial activity of cryptdin-2 against selected pathogens under physiological conditions. J Gastrointest Infect 2012; 2 (1):30-37.

DOI: 10.5005/jogi-2-1-30

License: CC BY-SA 4.0

Published Online: 01-06-2012

Copyright Statement:  Copyright © 2012; Jaypee Brothers Medical Publishers (P) Ltd.


Background and Objective: An initial step prior to clinical development of any therapeutically active peptide is to evaluate its stability under physiological conditions. As cationic antimicrobial peptides have been reported to lose their activity under physiological conditions, present study was done to evaluate the stability of antimicrobial activity of cryptdin-2 (a Paneth cell antimicrobial peptide) against Salmonella Typhimurium, Yersinia enterocolitica and Staphylococcus aureus in the presence and absence of physiological concentrations of bile salts, monovalent and divalent cations, trypsin as well as at various p H values. Methods: The antimicrobial activity of cryptdin-2 under various physiological conditions against Salmonella Typhimurium NCTC 74, Yersinia enterocolitica and Staphylococcus aureus was evaluated by use of a modified broth dilution technique Results: Interestingly, the activity of the peptide against the Gramnegative strainswas augmented by bile salts while no change in the activity against S. aureus was observed. Though there was a decrease in activity with increasing concentrations of metal ions, the activity was not completely lost. The peptide was able to sustain its activity against all the three test strains at physiological concentrations of trypsin. At p H8, no change in activitywas observed against Y. enterocolitica and S. Typhimurium while it was found to be reduced against S. aureus. Interpretation and Conclusion: The study provides data showing the stability of the peptide under the physiological conditions and indicates towards the possibility of developing it as an alternate strategy to combat bacterial pathogens.

PDF Share
  1. Liu SP, Zhou L, Lakshminarayanan R, Beuerman RW. Multivalent antimicrobial peptides as therapeutics: Design, principles and structural diversities. Int J Pept Res Ther 2010; 16:199–213.
  2. Dempsey CE, Hawrani A, Howe RA, Walsh TR. Amphipathic antimicrobial peptides—frombiophysics to therapeutics. Protein Pept Lett 2010;17:1334-44.
  3. Mc Cormick TS, Weinberg A. Epithelial cell-derived antimicrobial peptides are multifunctional agents that bridge innate and adaptive immunity. Periodontol 2000.2010;54:195– 206.
  4. Sharma S, Verma I, Khuller GK. Therapeutic potential of human neutrophil peptide-1 against experimental tuberculosis. Antimicrob Agents Chemother 2001;45:639–40.
  5. Rodríguez TM. On the cusp of change: new therapeutic modalities for HCV. Ann Hepatol 2010;9:123-31.
  6. Doss M, White MR, Tecle T, Gantz D, Crouch EC, Jung G et al. Interactions of alpha-, beta-, and theta-defensins with influenza A virus and surfactant protein D. J Immunol 2009;182:7878–87.
  7. Ouellette AJ, Hsieh MM, Nosek MT, Cano-Gauci DF, Huttner KM, Buick RN et al. Mouse Paneth cell defensins: primary structures and antibacterial activities of numerous cryptdin isoforms. Infect Immun 1994;62:5040–7.
  8. Preet S, Verma I, Rishi P. Cryptdin-2: a novel therapeutic agent for experimental Salmonella Typhimurium infection. J Antimicrob Chemother 2010;65:991-4.
  9. Piddock LJV, Ricci V, Mc Laren I, Griggs DJ. Role ofmutation in the gyr A and par C genes of nalidixic-acid-resistant Salmonella serotypes isolated from animals in the United Kingdom. J Antimicrob Chemother 1998;41:635–41.
  10. Rishi P, Mavi S, Bharrhan S, Shukla G, Tewari R. Protective efficacy of probiotic alone or in conjunction with a prebiotic in Salmonella-induced liver damage. FEMS Microbiol Ecol 2009;69:222–30.
  11. Porter EM, Dam EV, Valore EV, Ganz T. Broad-spectrum antimicrobial activity of human intestinal defensin 5. Infect Immun 1997;65:2396–401.
  12. Ouhara K, Komatsuzawa H, Yamada S, Shibra H, Fujiwara T, Ohara M et al. Susceptibilities of periodontopathogenic and cariogenic bacteria to antibacterial peptides, â-defensins and LL37, produced by human epithelial cells. J Antimicrob Chemother 2005;55:888–96.
  13. Wei GX, Campagna AN, Bobek LA. Factors affecting antimicrobial activity of MUC7 12-mer, a human salivarymucinderived peptide. Ann Clin Microbiol Antimicrob 2007; 6:14.doi:10.1186/1476-0711-6-14
  14. Preet S, Rishi P. Antimicrobial activity of Paneth cell derived cryptdin-2 against selected pathogens. Am J Biomed Sci 2010;2:13-22.
  15. Selsted ME, Miller SI, Henschen AH, Ouellette AJ. Enteric defensins: antibiotic peptide components of intestinal host defence. J Cell Biol 1992;118:929–36.
  16. Darkoh C, Lichtenberger LM, Ajami N, Dial EJ, Jiang ZD, Dupont HL et al. Bile acids improve the antimicrobial effect of rifaximin. Antimicrob Agents Chemother 2010;54: 3618-24.
  17. Wu G, Ding J, Li H, Li L, Zhao R, Shen Z, et al. Effects of cations and p H on antimicrobial activity of thanatin and sthanatin against Escherichia coli ATCC 25922 and B. subtilis ATCC 21332. Curr Microbiol 2008;57:552–7.
  18. Tomita T, Hitomi S, Nagase T, Matsui H, Matsuse T, Kimura S. Effect of ions on antibacterial activity of human beta defensin-2. Microbiol Immunol 2010;44:749–54.
  19. Hoover DM, Wu Z, Tucker K, Lu W, Lubkowski J. Antimicrobial characterization of human beta-defensin-3 derivatives. Antimicrob Agents Chemother 2003;47:2804–9.
  20. Bellamy W, Takase M, Wakabayashi H, Kawase K, Tomita M. Antibacterial spectrum of lactoferricin-B, a potent bactericidal peptide derived fromthe N-terminal region of bovine lactoferrin. J Appl Bacteriol 1992;73:472–9.
  21. Helmerhorst EJ, Breeuwer P, Hof VW, Walgreen WE, Oomen LC, Veerman EC. The cellular target of histatin-5 on Candida albicans is the energized mitochondrion. J Biol Chem 1999;274:7286–91.
  22. Cox DL, Sun Y, Liu H, Lehrer RI, Shafer WM. Susceptibility of Treponema pallidum to host-derived antimicrobial peptides. Peptides 2003;24:1741–6.
  23. Miyasaki KT, Iofel R, Oren A, Hyunh T, Lehrer RI. Killing of Fusobacterium nucleatum, Porphyromonas gingivalis and Prevotella intermedia by protegrins. J Periodontal Res 1998;33:91–8.
  24. Cole AM, Darouiche RO, Legarda D, Connell N, Diamond G. Characterization of a fish antimicrobial peptide: gene expression, subcellular localization, and spectrum of activity. Antimicrob Agents Chemother 2000;44:2039–45.
  25. Maisetta G, Luca MD, Esin S, Florio W, Brancatisano RL, Bottai D. Evaluation of the inhibitory effects of human serum components on bactericidal activity of human beta defensin-3. Peptides 2008;29:1-6.
  26. Evans DF, Pye G, Bramley R, Clark AG, Dyson TJ, Hardcastle JD, et al. Measurement of gastrointestinal p H profiles in normal ambulant human subjects. Gut 1988;29:1035–41.
  27. Fallingborg J, Christensen LA, Nielsen MI, Jacopson BA, Abildquard K, Rasmussen HH et al. Measurement of gastrointestinal p Hand regional transit times in normal children. J Pediatr Gastroenterol Nutr 1990;11:211–4.
  28. Mc Cloy RF, Greenberg GR, Baron JH. Duodenal p H in health and duodenal ulcer disease: effect of ameal, coca-cola, smoking, and cimetidine. Gut 1984;25:386–92.
  29. Shimoda M, Ohki K, Shimamoto Y, Kohashi O. Morphology of defensin-treated Staphylococcus aureus. Infect Immun 1995;63:2886–91.
  30. Peschel A, Jack RW, Otto M, Collins LV, Staubitz P, Nicholson G et al. Staphylococcus aureus resistance to human defensins and evasion of neutrophil killing via the novel virulence factor mpr F is based onmodification of membrane lipidswith L-lysine. J Exp Med 2001;193:1067-76.
  31. Peschel A, Otto M, Jack RW, Kalbacher H, Jung G, Gotz F et al. Inactivation of the dlt operon in Staphylococcus aureus confers sensitivity to defensins, protegrins, and other antimicrobial peptides. J Biol Chem 1999;274:8405-510.
  32. Selsted ME, Szklarek D, Ganz T, Lehror RI. Activity of rabbit leukocyte peptides against Candida albicans. Infect Immun 1985;49:202–6.
  33. Bessalle R, Haas H, Goria A, Shalit I, Fridkin M. Augmentation of the antibacterial activity of magainin by positive-charge chain extension. Antimicrob Agents Chemother 1992;36: 313-7.
  34. Eby DM, Farrington KE, Johnson GR. Synthesis of bioinorganic antimicrobial peptide nanoparticles with potential therapeutic properties. Biomacromolecules 2008;9:2487-94.
PDF Share
PDF Share

© Jaypee Brothers Medical Publishers (P) LTD.