In vivo efficiency of the produced recombinant lysostaphin antimicrobial peptide in treatment of methicillin-resistant Staphylococcus aureus (MRSA) skin infection in a mouse model
Abstract
Background and Objectives: Staphylococcus simulans secretes an antimicrobial compound called lysostaphin, which has bactericidal properties. It destroys staphylococci through the hydrolysis of peptidoglycan in the cell wall. Therefore, this unique property indicates the high ability of lysostaphin in the treatment of staphylococcal infections and is considered as an anti-staphylococcal agent.
Materials and Methods: Escherichia coli BL21 (DE3) competent cells were transformed with pET32a-lysostaphin clone and induced by isopropyl-β-D-thio-galactoside (IPTG). The recombinant protein was purified by affinity chromatography. Recombinant lysostaphin -A-based ointment was used for external wound healing in animal model. In vivo activity of ointment was evaluated by clinical evidences and cytological microscopic assessment.
Results: Our results showed the recombinant protein was produced exactly. The results of checkerboard tests showed MIC, MBC and antibacterial activity test an acute reduction of cell viability during the use of lysostaphin, and SEM results approved the intense wrecking effects of lysostaphin in combination on bacterial cells. Macroscopic findings and microscopic data showed that the recombinant lysostaphin ointment was effective on excisional wound healing.
Conclusion: Our findings proved that the recombinant lysostaphin ointment was effective on wound healing due to Staphylococcus aureus infection.
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14. CLSI Document M07-A10. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically; Clinical and Laboratory Standards Institute: Wayne, PA, USA, 2015.
15. 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 thymol. Molecules 2017; 22: 822.
16. Schaffer AC, Solinga RM, Cocchiaro J, Portoles M, Kiser KB, Risley A, et al. Immunization with Staphylococcus aureus clumping factor B,a major determinant in nasal carriage, reduces nasal colonization in a murine modle. Infect Immun 2006; 74: 2145-2153.
17. Liu Y, Lim J, Teoh SH. Review: Development of clinically relevant scaff; olds for vascularised bone tissue engineering. Biotechnol Adv 2013; 31: 688-705.
18. 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.
19. Bastos MD, Coutinho BG, Coelho ML. Lysostaphin: A Staphylococcal bacteriolysin with potential clinical applications. Pharmaceuticals (Basel) 2010; 3: 1139-1161.
20. Jayakumar J, Kumar VA, Biswas L, Biswas R. Therapeutic applications of lysostaphin against Staphylococcus aureus. J Appl Microbiol 2021; 131: 1072-1082.
21. Kokai-Kun JF, Chanturiya T, Mond JJ. Lysostaphin as a treatment for systemic Staphylococcus aureus infection in a mouse model. J Antimicrob Chemother 2007; 60: 1051-1059.
22. Kokai-Kun JF, Walsh SM, Chanturiya T, Mond JJ. Lysostaphin cream eradicates Staphylococcus aureus nasal colonization in a cotton rat model. Antimicrob Agents Chemother 2003; 47: 1589-1597.
23. Grüdling A, Schneewind O. Cross-linked peptidoglycan mediates lysostaphin binding to the cellwall envelope of Staphylococcus aureus. J Bacteriol 2006; 188: 2463-2472.
24. Satishkumar R, Sankar S, Yurko Y, Lincort A, Shipp J, Heniford BT, et al. Evaluation of the antimicrobial activity of lysostaphin-coated hernia repair meshes. Antimicrob Agents Chemother 2011; 55: 4379-4385.
25. Dixon RE, Goodman JS, Koenig MG. Lysostaphin: an enzymatic approach to staphylococcaldisease. III. Combined lysostaphin-methicillin therapy of established staphylococcal abscesses inmice. Yale J Biol Med 1968; 41: 62-68.
26. Patron RL, Climo MW, Goldstein BP, Archer GL. Lysostaphin treatment of experimentalaortic valve endocarditis caused by a Staphylococcus aureus isolate with reduced susceptibility tovancomycin. Antimicrob Agents Chemother 1999; 43: 1754-1755.
2. Mistry RD. Skin and soft tissue infections. Pediatr Clin North Am 2013; 60: 1063-1082.
3. Haq FU, Imran M, Saleem S, Aftab U, Ghazal A. Investigation of Morchella esculenta and Morchella conica for their antibacterial potential against methicillin-susceptible Staphylococcus aureus, methicillin-resistant Staphylococcus aureus and Streptococcus pyogenes. Arch Microbiol 2022; 204: 391.
4. Lakhundi S, Zhang K. Methicillin-resistant Staphylococcus aureus: molecular characterization, evolution, and epidemiology. Clin Microbiol Rev 2018; 31(4): e00020-18.
5. Koehl JL, Muthaiyan A, Jayaswal RK, Ehlert K, Labischinski H, Wilkinson BJ. Cell wall composition and decreased autolytic activity and lysostaphin susceptibility of glycopeptide-intermediate Staphylococcus aureus. Antimicrob Agents Chemother 2004; 48: 3749-3757.
6. Bastos MdCdF, Coutinho BG, Coelho MLV. Lysostaphin: a staphylococcal bacteriolysin with potential clinical applications. Pharmaceuticals (Basel) 2010; 3: 1139-1161.
7. Garde S, Chodisetti PK, Reddy M. Peptidoglycan: structure, synthesis, and regulation. EcoSal Plus 2021; 9: 10.1128/ecosalplus.
8. Dajcs JJ, Hume EB, Moreau JM, Caballero AR, Cannon BM,O’Callaghan RJ. Lysostaphin treatment of methicillin-resistant Staphylococcus aureus keratitis in the rabbit. Invest Ophthalmol Vis Sci 2000; 41: 1432-1437.
9. Kumar JK. Lysostaphin: an antistaphylococcal agent. Appl Microbiol Biotechnol 2008; 80: 555-561.
10. Bastos MC, Ceotto H, Coelho ML, Nascimento JS. Staphylococcal antimicrobial peptides: relevant properties and potential biotechnological applications. Curr Pharm Biotechnol 2009; 10: 38-61.
11. Farhangnia L, Ghaznavi- Rad E, Mollaee N, Abtahi H. Cloning, expression, and purification of recombinant lysostaphin from Staphylococcus simulans. Jundishapur J Microbiol 2014; 7(5): e10009.
12. Mahmoudi S, Abtahi H, Bahador A, Mosayebi G, Salmanian AH. Production of recombinant streptokinase in E. coli and reactivity with immunized mice. Pak J Biol Sci 2010; 13: 380-384.
13. 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.
14. CLSI Document M07-A10. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically; Clinical and Laboratory Standards Institute: Wayne, PA, USA, 2015.
15. 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 thymol. Molecules 2017; 22: 822.
16. Schaffer AC, Solinga RM, Cocchiaro J, Portoles M, Kiser KB, Risley A, et al. Immunization with Staphylococcus aureus clumping factor B,a major determinant in nasal carriage, reduces nasal colonization in a murine modle. Infect Immun 2006; 74: 2145-2153.
17. Liu Y, Lim J, Teoh SH. Review: Development of clinically relevant scaff; olds for vascularised bone tissue engineering. Biotechnol Adv 2013; 31: 688-705.
18. 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.
19. Bastos MD, Coutinho BG, Coelho ML. Lysostaphin: A Staphylococcal bacteriolysin with potential clinical applications. Pharmaceuticals (Basel) 2010; 3: 1139-1161.
20. Jayakumar J, Kumar VA, Biswas L, Biswas R. Therapeutic applications of lysostaphin against Staphylococcus aureus. J Appl Microbiol 2021; 131: 1072-1082.
21. Kokai-Kun JF, Chanturiya T, Mond JJ. Lysostaphin as a treatment for systemic Staphylococcus aureus infection in a mouse model. J Antimicrob Chemother 2007; 60: 1051-1059.
22. Kokai-Kun JF, Walsh SM, Chanturiya T, Mond JJ. Lysostaphin cream eradicates Staphylococcus aureus nasal colonization in a cotton rat model. Antimicrob Agents Chemother 2003; 47: 1589-1597.
23. Grüdling A, Schneewind O. Cross-linked peptidoglycan mediates lysostaphin binding to the cellwall envelope of Staphylococcus aureus. J Bacteriol 2006; 188: 2463-2472.
24. Satishkumar R, Sankar S, Yurko Y, Lincort A, Shipp J, Heniford BT, et al. Evaluation of the antimicrobial activity of lysostaphin-coated hernia repair meshes. Antimicrob Agents Chemother 2011; 55: 4379-4385.
25. Dixon RE, Goodman JS, Koenig MG. Lysostaphin: an enzymatic approach to staphylococcaldisease. III. Combined lysostaphin-methicillin therapy of established staphylococcal abscesses inmice. Yale J Biol Med 1968; 41: 62-68.
26. Patron RL, Climo MW, Goldstein BP, Archer GL. Lysostaphin treatment of experimentalaortic valve endocarditis caused by a Staphylococcus aureus isolate with reduced susceptibility tovancomycin. Antimicrob Agents Chemother 1999; 43: 1754-1755.
| Files | ||
| Issue | Vol 15 No 2 (2023) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v15i2.12476 | |
| Keywords | ||
| Anti-bacterial agents; Lysostaphin; Staphylococcus aureus; Wound healing | ||
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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Sadoogh Abbasian S, Ghaznavi-Rad E, Sadelaji S, Abtahi H. In vivo efficiency of the produced recombinant lysostaphin antimicrobial peptide in treatment of methicillin-resistant Staphylococcus aureus (MRSA) skin infection in a mouse model. Iran J Microbiol. 2023;15(2):243-250.
