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Dispersal of Pseudomonas aeruginosa biofilm using TDS97 bacteriophage and recombinant alginate lyase enzyme

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Abstract

Pseudomonas aeruginosa is an opportunistic human pathogen capable of forming biofilms and contaminate medical devices and food. Bacteremia caused by this organism is one of the most serious complications with a mortality rate from 18 to 61%. This study aimed investigate the synergistic effect between recombinant alginate lyase enzyme and bacteriophage TDS 97 in controlling P. aeruginosa biofilm. TDS97 bacteriophage was isolated from municipal wastewater and purified by the Top-Agar method. Electron microscopy revealed that this phage belonged to the Myoviridae family. Evaluation of the effect of phage on P. aeruginosa biofilm showed that phage significantly inhibited both biofilm formation (about 82%) and was able to cause dispersal of biofilm (79%). Alginate lyase enzyme was purified at a concentration of 130 μg/ml. Its antibacterial effects on P. aeruginosa were investigated. The results showed that the enzyme had a significant effect on inhibiting biofilm formation (about 74%) and its dispersion (about 71%). The synergistic effect of phage and enzyme was evaluated and the results showed that these two anti-biofilm effects strengthen each other (> 81% inhibition, > 77% dispersion). The results of this study demonstrated that the use of recombinant alginate lyase enzyme with bacteriophage TDS 97 may provide a suitable therapeutic alternative for controlling P. aeruginosa biofilm.

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All data generated or analyzed during this study are included in this published article.

Abbreviations

EPS:

Extracellular Polymeric Substances

eDNA:

Extracellular DNA

LB:

Luria-Bertani

SMB:

Sodium chloride, Magnesium sulfate buffer

TSB:

Tryptic Soy Broth

PBS:

Phosphate-Buffered Saline

FBIC:

Fractional index of Biofilm formation Inhibitor Concentration

FBEC:

Fractional index of Biofilm Eradication Concentration

MOI:

Multiplicity of Infection

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Acknowledgements

The authors would like to acknowledge Dr. Jalalei (Isfahan University of Technology, Iran) for providing the vector pET-28a, and the staff of bacteriology and biotechnology laboratories (Faculty of Veterinary Medicine, Department of Pathobiology, Shiraz, Iran) for providing us technical supports.

Funding

This work was funded by Shiraz University (Grant No. 9430195).

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Authors and Affiliations

  1. Department of Pathobiology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran

    Leila Dolatshah, Mohammad Tabatabaei & Mohammad Sadeghpour Mobarakeh

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  1. Leila Dolatshah
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Contributions

This study is the major component of the work toward the PhD thesis of the LD supervised by MT. LD conceived and performed the experiments; analyzed and interpreted the data. MS collaborated in bacteriophage isolation and also confirmed the diagnosis of Pseudomonas aeruginosa isolates. LD wrote the first draft of the manuscript. Then, the whole manuscript critically revised by MT. MT helped in planning the project; contributed to revision of the manuscript; supervised the project. All authors read, edited, and approved the final version of the manuscript.

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Correspondence to Mohammad Tabatabaei.

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Dolatshah, L., Tabatabaei, M. & Sadeghpour Mobarakeh, M. Dispersal of Pseudomonas aeruginosa biofilm using TDS97 bacteriophage and recombinant alginate lyase enzyme. J Proteins Proteom (2024). https://doi.org/10.1007/s42485-024-00154-8

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