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Investigation of the antibacterial activity of a short cationic peptide against multidrug-resistant Klebsiella pneumoniae and Salmonella typhimurium strains and its cytotoxicity on eukaryotic cells

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Abstract

With the growing microbial resistance to conventional antimicrobial agents, the development of novel and alternative therapeutic strategies are vital. During recent years novel peptide antibiotics with broad spectrum activity against many Gram-positive and Gram-negative bacteria have been developed. In this study, antibacterial activity of CM11 peptide (WKLFKKILKVL-NH2), a short cecropin–melittin hybrid peptide, is evaluated against antibiotic-resistant strains of Klebsiella pneumoniae and Salmonella typhimurium as two important pathogenic bacteria. To appraise the antibacterial activity, minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC) and bactericidal killing assay were utilized with different concentrations (2–128 mg/L) of peptide. To evaluate cytotoxic effect of peptide, viability of RAJI, Hela, SP2/0, CHO, LNCAP cell lines and primary murine macrophage cells were also investigated with MTT assay in different concentrations (3–24 and 0.5–16 mg/L, respectively). MICs of K. pneumoniae and S. typhimurium isolates were in range of 8–16 and 4–16 mg/L, respectively. In bactericidal killing assay no colonies were observed at 2X MIC for K. pneumoniae and S. typhimurium isolates after 80–90 min, respectively. Despite the fact that CM11 reveals no significant cytotoxicity on RAJI, Hela, SP2/0, and CHO cell lines beneath 6 mg/L at first 24 and 48 h, the viability of LNCAP cells are about 50 % at 3 mg/L, which indicates strong cytotoxicity of the peptide. In addition, macrophage toxicity by MTT assay showed that LD50 of CM11 peptide is 12 μM (16 mg/L) after 48 h while in this concentration after 24 h macrophage viability was about 70 %.

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Acknowledgments

This study was supported and conducted by Applied Biotechnology Research Center. The authors like to thanks Dr. Ali Mohammad Latifi for his kind and generous assistance.

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Authors and Affiliations

  1. Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, P.O. Box 19395-5487, Tehran, Iran

    Mehrdad Moosazadeh Moghaddam & Jafar Amani

  2. Department of Molecular Biology, Faculty of Sciences, University of Mohaghegh Ardabili, Ardabil, Iran

    Kamal Azizi Barjini

  3. Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

    Mahdi Fasihi Ramandi

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  1. Mehrdad Moosazadeh Moghaddam
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Correspondence to Mehrdad Moosazadeh Moghaddam.

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Moghaddam, M.M., Barjini, K.A., Ramandi, M.F. et al. Investigation of the antibacterial activity of a short cationic peptide against multidrug-resistant Klebsiella pneumoniae and Salmonella typhimurium strains and its cytotoxicity on eukaryotic cells. World J Microbiol Biotechnol 30, 1533–1540 (2014). https://doi.org/10.1007/s11274-013-1575-y

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