The Antimicrobial Effects of TP4-LYC1 Fusion Protein against Multidrug-Resistant A. Baumannii

Document Type : Original Article (s)

Authors

1 Professor, Department of Clinical Pharmacy and Pharmacy Practice, School of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran

2 Assistant Professor, Department of Pharmaceutical Biotechnology, School of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran

Abstract

Background: Colistin, tigecycline and ampicillin/sulbactam (high dose) are the only antibiotics with high efficacy against Acinetobacter baumannii making them the last line of treatment. So, the investigation of new antimicrobial agents with potential activity against these pathogens is crucial. Antimicrobial peptides are natural agents that play an important role in innate immunity against various types of pathogens. In this project, the antimicrobial activity of TP4-LYC1, an antimicrobial peptide consisting of Tilapia Piscidin 4 (TP4) with lycosin-1, was investigated against resistant A. baumannii as well as the synergistic effects of this chimeric peptide with colistin and meropenem.
Methods: Optimization of the cleavage process for each peptides (TP4, LYC1, and TP4-LYC1) to obtain the maximum amount of peptides was performed based on the incubation temperature, time of incubation and the cleavage buffer pH. Finally, the antimicrobial effects of each peptide, alone or in combination with colistin or meropenem were analyzed by broth micro-dillusion and checker board methods, respectively.
Findings: The best conditions for cleaving TP4, were pH 6.5 with a 24 Hours incubation time. These conditions for LYC1 and the chimeric peptide were concluded as pH 4 with incubation time of 24 and 48 Hours, respectively. The antibacterial activity of TP4 was lower than TP4-LYC1 for each biological sample. Also, TP4-LYC1 exhibited synergistic effects with Meropenem and Colistin in most cases.
Conclusion: TP4-LYC1 shows potential as a candidate for the treatment of infections caused by A. baumanni in combination with routine effective antibiotics in order to reduce the dosage of these antibiotics.

Keywords

Main Subjects

References

  1. Gröschel MI, Meehan CJ, Barilar I, Diricks M, Gonzaga A, Steglich M, et al. The phylogenetic landscape and nosocomial spread of the multidrug-resistant opportunist Stenotrophomonas maltophilia. Nat Commun 2020; 11(1): 2044.
  2. Giraldi G, Montesano M, Napoli C, Frati P, La Russa R, Santurro A, et al. Healthcare-associated infections due to multidrug-resistant organisms: a surveillance study on extra hospital stay and direct costs. Curr Pharm Biotechnol 2019; 20(8): 643-52.
  3. Sobouti B, Mirshekar M, Fallah S, Tabaei A, Mehrabadi JF, Darbandi A. Pan drug-resistant Acinetobacter baumannii causing nosocomial infections among burnt children. Med J Islam Repub Iran 2020; 34: 24.
  4. Li X, Song Y, Wang L, Kang G, Wang P, Yin H, et al. A potential combination therapy of berberine hydrochloride with antibiotics against multidrug-resistant Acinetobacter baumannii. Front Cell Infect Microbiol 2021; 11: 660431.
  5. Huan Y, Kong Q, Mou H, Yi H. Antimicrobial peptides: classification, design, application and research progress in multiple fields. Front Microbiol 2020; 11: 582779.
  6. Neshani A, Akbari Eidgahi MR, Zare H, Ghazvini K. Extended-Spectrum antimicrobial activity of the Low cost produced Tilapia Piscidin 4 (TP4) marine antimicrobial peptide. J Res Med Dent Sci 2018; 6(5): 327-4.
  7. Pan CY, Chen JC, Chen TL, Wu JL, Hui CF, Chen JY. Piscidin is highly active against carbapenem-resistant Acinetobacter baumannii and NDM-1-producing Klebsiella pneumonia in a systemic septicaemia infection mouse model. Mar Drugs 2015; 13(4): 2287-305.
  8. Liu Z, Deng M, Xiang J, Ma H, Hu W, Zhao Y, et al. A novel spider peptide toxin suppresses tumor growth through dual signaling pathways. Curr Mol Med 2012; 12(10): 1350-60.
  9. Tan H, Ding X, Meng S, Liu C, Wang H, Xia L, et al.
    Antimicrobial potential of lycosin-I, a cationic and amphiphilic peptide from the venom of the spider Lycosa singorensis. Curr Mol Med 2013; 13(6): 900-10.
  10. Wang L, Wang YJ, Liu YY, Li H, Guo LX, Liu ZH,
    et al. In vitro potential of lycosin-I as an alternative antimicrobial drug for treatment of multidrug-resistant Acinetobacter baumannii infections. Antimicrob Agents Chemother 2014; 58(11): 6999-7002.
  11. Sambrook J, Russell DW. The condensed protocols from molecular cloning: a laboratory manual. New York, NY: CSHL Press; 2006.
  12. New England Biolabs. IMPACTTM-TWIN. Purification, ligation and cyclization of recombinant proteins usinf self-cleavable affinity tag. version 1.5, 7/08. [Online]. Avilable from: URL:

https://www.neb.com/-/media/nebus/files/manuals/impact-twin-manual-e6950-708.pdf?rev=b5d93033cb034cba9987c836e30d3c4e&hash=1D202C22004B16D256C88047FAB2A3D5

  1. Kruger NJ. The Bradford method for protein quantitation. In: Kruger NJ, editor. The protein protocols handbook. Berlin, Heidelberg: Springer; 2009. p. 17-24.
  2. Safarika A, Galani I, Pistiki A, Giamarellos-Bourboulis EJ. Time-kill effect of levofloxacin on multidrug-resistant Pseudomonas aeruginosa and Acinetobacter baumannii: synergism with imipenem and colistin. Eur J Clin Microbiol Infect Dis 2015; 34(2): 317-23.
  3. Li Y. Self-cleaving fusion tags for recombinant protein production. Biotechnol Lett 2011; 33(5): 869-81.
  4. Mujika JI, Lopez X, Mulholland AJ. Mechanism of C-terminal intein cleavage in protein splicing from QM/MM molecular dynamics simulations. Organic Biomol Chem 2012; 10(6): 1207-18.
  5. Shafiee F, Minaiyan G, Moazen F. Jahanian-najafabadi A. Recombinant production and intein-mediated purification of an antimicrobial peptide, BR2. Int J Pept Res Ther 2017; 23: 501-7.
  6. Hou J, Liu HY, Diao H, Yu H. The truncated human beta-defensin 118 can modulate lipopolysaccharide mediated inflammatory response in RAW264.7 macrophages. Peptides 2021; 136: 170438.
  7. Xing L, Wu W, Zhou B, Lin Z. Streamlined protein expression and purification using cleavable self-aggregating tags. Microb Cell Fact 2011; 10(1): 42.
  8. Su BC, Liu YC, Ting CH, Lyu PC, Chen JY. Antimicrobial peptide TP4 targets mitochondrial adenine nucleotide translocator 2. Mar Drugs 2020; 18(8): 417.‏
  9. Pan CY, Tsai TY, Su BC, Hui CF, Chen JY. Study of the antimicrobial activity of tilapia piscidin 3 (TP3) and TP4 and their effects on immune functions in hybrid tilapia (Oreochromis spp.). PloS One 2017; 12(1): e0169678.
  10. Pan CY, Chen JC, Chen TL, Wu JL, Hui CF, Chen JY. Piscidin is highly active against carbapenem-resistant Acinetobacter baumannii and NDM-1-producing Klebsiella pneumonia in a systemic septicaemia infection mouse model. Mar Drugs 2015; 13(4): 2287-305.
  11. Tan H, Ding X, Meng S, Liu C, Wang H, Xia L, et al. Antimicrobial potential of lycosin-I, a cationic and amphiphilic peptide from the venom of the spider Lycosa singorensis. Curr Mol Med 2013; 13(6): 900-10.

 

Journal of Isfahan Medical School
Volume 41, Issue 729
1st Week, October
September and October 2023
Pages 615-623

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