Ib-AMP4 antimicrobial peptide as a treatment for skin and systematic infection of methicillin-resistant Staphylococcus aureus (MRSA)

Document Type : Original Article

Authors

1 Molecular and Medical Research Center, Arak University of Medical Sciences, Arak, Iran

2 Department of Medical Laboratory Sciences, Arak School of Paramedicine, Arak University of Medical Sciences, Arak, Iran

Abstract

Objective(s): Antimicrobial peptide compounds (AMPs) play important roles in the immune system. They also exhibit significant anti-tumor and antibacterial properties. Most AMPs are cationic and are able to bind bacterial cell membranes through electrostatic affinity. Ib-AMP4 is a plant-derived AMP that exerts rapid bactericidal functions. In the present study, the antibacterial efficiency of the produced recombinant Ib-AMP4 in elimination of Methicillin-resistant Staphylococcus aureus (MRSA) bacterial infection, was investigated under in vitro and in vivo situations. 
Materials and Methods: The synthesized Escherichia coli codon-optimized gene sequences of the Ib-AMP4 were expressed in E. coli BL21 (DE3) pLysS. The recombinant Ib-AMP4 was purified and refolding conditions were optimized. The antibacterial efficiency of the refolded peptide against MRSA was tested under in vivo and in vitro situations for treatment of skin and systematic infection of MRSA in a mouse model.
Results: Antibacterial assays confirmed the antibacterial function of Ib-AMP4 against MRSA. SEM results proved the destructive effects of applying Ib-AMP4 on MRSA biomembrane. Time-kill curve and growth kinetic assay illustrated rapid antibacterial activity of the produced Ib-AMP4. Moreover, Ib-AMP4 showed significant infection treatment ability in a mouse model and all infected mice receiving Ib-AMP4 protein survived and there was no trace of bacteria in their blood samples.
Conclusion: The results confirmed the rapid antibacterial potential of the produced recombinant Ib-AMP4 to be used for efficient treatment of  MRSA infection.

Keywords

References

1. Avison MB. New approaches to combating antimicrobial drug resistance. Genome Biol 2005;6:243-246.
2. Sarkar T, Chetia M & Chatterjee, S. Antimicrobial peptides and proteins: From nature’s reservoir to the laboratory and beyond. Front Chem 2021;9:691532. 
3. Mukhopadhyay S, Bharath Prasad AS, Mehta CH & Nayak UY. Antimicrobial peptide polymers: No escape to ESKAPE pathogens-a review. World J Microbiol Biotechnol 2020;36:131-144.
4. Flórez-Castillo JM, Ropero-Vega JL, Perullini M, & Jobbágy M. Biopolymeric pellets of polyvinyl alcohol and alginate for the encapsulation of Ib-M6 peptide and its antimicrobial activity against Escherichia coli. Heliyon 2019; 5: e01872.
5. Sadoogh Abbasian S, Abtahi H, Zolfaghari MR, Soufian S and Ghaznavi-Rad E. Cloning, expression,purification and antigenic evaluation of hyaluronidase antigenic fragments recombinant protein of streptococcus pyogenes . Afr J Biotechnol 2012;11:2376-2380.
6. Mirjamali N, Soufian, S, Molaee N, Sadoogh Abbasian S, Abtahi H. Cloning and expression of the enzymatic region of Streptococcal hyaluronidase. Iran J Basic Med Sci 2014; 17:667–672.
7. Abtahi H, Salmanian AH, Rafati S, Nejad GB, Hassan ZM. High level expression of recombinant ribosomal protein (L7/L12) from Brucella abortus and its reaction with infected human sera. Iran Biomed J 2004; 8:13–18.
8. Sadoogh Abbasian S, Ghaznavi-Rad E, Akbari N, Zolfaghari M R, pakzad I and Abtahi H. Overexpression and enzymatic assessment of antigenic fragments of hyaluronidase recombinant protein from streptococcus pyogenes. Jundishapur J Microbiol 2015; 8: e13653.
9. Fahimirad S, Razavi SH, Abtahi H, Alizadeh H, Ghorbanpour M. Recombinant production and antimicrobial assessment of beta casein-IbAMP 4 as a novel antimicrobial polymeric protein and its synergistic effects with thymol. Int J Pept Res Ther 2018;24:213-222.
10. Sadoogh Abbasian S, Soufian S, Ghaznavi-Rad E, Abtahi H. High level activity of recombinant lysostaphin after computer simulation and additive-based refolding. Int J Pept Res Ther 2019; 25:1241–1249.
11. Fahimirad S, Abtahi H, Razavi SH, Alizadeh H, Ghorbanpour M. Production of recombinant antimicrobial polymeric protein beta casein-E 50-52 and its antimicrobial synergistic effects assessment with yhymol. Molecules 2017;22:822-836.
12. Elshikh M, Ahmed S, Funston S, Dunlop P, McGaw M, Marchant R, et al. Resazurin-based 96-well plate microdilution method for the determination of minimum inhibitory concentration of biosurfactants. Biotechnol Lett 2016;38:1015-1019.
13. Fahimirad S, Ghaznavi-Rad E, Abtahi H, Sarlak N. Antimicrobial activity, stability and wound healing performances of chitosan nanoparticles loaded recombinant LL37 antimicrobial peptide. Int J Pept Res Ther 2021;27:2505-2515.
14. Rudilla H, Fusté E, Cajal Y, Rabanal F, Vinuesa T, Viñas M. Synergistic antipseudomonal effects of synthetic peptide AMP38 and carbapenems. Molecules 2016;21:1223-1234.
15. Zengin H, Baysal AH. Antibacterial and antioxidant activity of essential oil terpenes against pathogenicand spoilage-forming bacteria and cell structure-activity relationships evaluated by SEM
microscopy . Molecules 2014;19:17773-17798.
16. Soković M, Glamočlija J, Marin PD, Brkić D, van Griensven LJ. Antibacterial effects of the essential oils of commonly consumed medicinal herbs using an in vitro model. Molecules 2010;15:7532-7546. 
17. Cirioni O, Giacometti A, Ghiselli R, Bergnach C, Orlando F, Silvestri C, et al. LL-37 protects rats against lethal sepsis caused by Gram-negative bacteria. Antimicrob Agents Chemother 2006;50:1672-1679.
18. Nguyen LT, Haney EF, Vogel HJ. The expanding scope of antimicrobial peptide structures and their modes of action. Trends Biotechnol 2011;29:464-472.
19. Pushpanathan M, Gunasekaran P, Rajendhran J. Antimicrobial peptides: versatile biological properties. Int J Pept 2013; 2013:675391.
20. Mann NH. The potential of phages to prevent MRSA infections. Res Microbiol 2008;159:400-405.
21. Fan X, Schäfer H, Reichling J, Wink M. Bactericidal properties of the antimicrobial peptide Ib-AMP4 from Impatiens balsamina produced as a recombinant fusion-protein in Escherichia coli.Biotechnol J 2013;8:1213-1220.
22. Satei P, Ghaznavi-Rad E, Fahimirad S, Abtahi H. Recombinant production of Trx-Ib-AMP4 and Trx-E50-52 antimicrobial peptides and antimicrobial synergistic assessment on the treatment of methicillin-resistant Staphylococcus aureus under in vitro and in vivo situations. Protein Expr Purif 2021;188:105949.
23. Fan X, Reichling J, Wink M. Antibacterial activity of the recombinant antimicrobial peptide Ib-AMP4 from Impatiens balsamina and its synergy with other antimicrobial agents against drug resistant bacteria. Pharmazie 2013; 68:628-630.
Iranian Journal of Basic Medical Sciences
Volume 25, Issue 2
February 2022
Pages 232-238

Files

Share

How to cite

Statistics