Probiotics and Antimicrobial Proteins

, Volume 11, Issue 3, pp 1034–1041 | Cite as

Secretory Expression of a Chimeric Peptide in Lactococcus lactis: Assessment of its Cytotoxic Activity and a Deep View on Its Interaction with Cell-Surface Glycosaminoglycans by Molecular Modeling

  • Abbas Tanhaeian
  • Mahmoud Reza Jaafari
  • Farajollah Shahriari Ahmadi
  • Roghayyeh Vakili‐Ghartavol
  • Mohammad Hadi SekhavatiEmail author


Nowadays, cancer remains a major cause of death affecting millions of people. Currently, the antimicrobial peptides (AMPs) as potent anticancer therapeutic agents offer specificity and low levels of side effects in cancer therapy. In the present study, a cationic chimeric peptide (cLFchimera), derived from camel lactoferrin, was expressed as a secretory peptide using P170 expression system in L. lactis. Peptide purification was carried out using Ni-NTA agarose column from culture medium with 21 μ/mL concentration. The recombinant peptide was investigated for its activity against four tumor and one normal cell line. The cLFchimera was more active against two tumor cell lines (chondrosarcoma and colorectal cancer cells), but the activity against two other tumor cell lines (hepatoma and breast cancer cell line) and normal cells was low. Finally, to have better insight into the mode of action of the peptide on cytotoxic activity, we examined the interaction of cationic peptide with two glycosaminoglycans (GAGs), heparan sulfate (HS) and chondroitin sulfate (CS), as the two most anionic molecules on the cell surface by molecular dynamic simulation. The results of in silico analysis showed that the cLFchimera interacted with HS and CS with a totally different amino acid profile. Hydrogen bonding screening in GAGs-peptide complexes revealed K21, V23 and I3, R16 are the dominant amino acids involved in peptide-HS and CS interaction, respectively. Overall, the results of this investigation showed the P170 expression system successfully expressed a cationic peptide with potent anticancer activity. Moreover, molecular docking analysis revealed the pattern of peptide interaction with negatively charged membrane molecules.


P170 expression system Lactococcus lactis Cytotoxic activity Glycosaminoglycans Molecular dynamic simulation 



We are very grateful to Miss Marjan Azghandi and Dr. Reza Majidzade for their outstanding technical assistance.

Authors’ Contributions

MHS and MRJ planed the experiment. Cloning, protein expression, and western blotting analysis were performed by AT and FSA. Cytotoxicity assay was carried out by RV. MHS and AB did the data collection. MHS performed MD simulation. MHS, AB, and MRJ interpreted the data. MHS prepared the manuscript.


This work was supported by Ferdowsi University of Mashhad, Iran with grant No.38736.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Abbas Tanhaeian
    • 1
  • Mahmoud Reza Jaafari
    • 2
    • 3
  • Farajollah Shahriari Ahmadi
    • 1
  • Roghayyeh Vakili‐Ghartavol
    • 4
  • Mohammad Hadi Sekhavati
    • 5
    Email author
  1. 1.Department of Biotechnology and Plant Breeding, Faculty of AgricultureFerdowsi University of MashhadMashhadIran
  2. 2.Nanotechnology Research Center, Pharmaceutical Technology InstituteMashhad University of Medical SciencesMashhadIran
  3. 3.Department of Pharmaceutical Nanotechnology, School of PharmacyMashhad University of Medical SciencesMashhadIran
  4. 4.Department of Medical Nanotechnology, School of Advanced Technologies in MedicineTehran University of Medical SciencesTehranIran
  5. 5.Department of Animal Sciences, Faculty of AgricultureFerdowsi University of MashhadMashhadIran

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