Analysis of antibacterial and antibiofilm activity of purified recombinant Azurin from Pseudomonas aeruginosa
| Autor: | Sharareh Moghim, Bahram Nasr-Esfahani, Bita Bakhshi, Hajar Mohammadi-Barzelighi, Hossein Fazeli, Bahram Daraei |
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| Rok vydání: | 2019 |
| Předmět: |
Microbiology (medical)
Antimicrobial peptides lcsh:QR1-502 Bacterial growth medicine.disease_cause 01 natural sciences Microbiology lcsh:Microbiology Shigella flexneri Affinity chromatography Recombinant Azurin 0502 economics and business medicine 0101 mathematics Escherichia coli biology Pseudomonas aeruginosa Chemistry 010102 general mathematics 05 social sciences biology.organism_classification Antibacterial effect Vibrio cholerae Antibiofilm activity Azurin 050203 business & management |
| Zdroj: | Iranian Journal of Microbiology, Vol 11, Iss 2 (2019) |
| ISSN: | 2008-4447 2008-3289 |
| Popis: | Background and Objectives: The aim of this study was to evaluate the antibacterial and antibiofilm activity of recombinant Azurin from Pseudomonas aeruginosa against different bacterial species. Materials and Methods: The azurin gene was cloned in the pET21a vector. The pET21a-azurin construct was transformed into Escherichia coli BL21. The recombinant Azurin was expressed and purified using affinity chromatography and con- firmed by Western blotting. The cytotoxicity of rAzurin was assessed on peripheral blood mononuclear cells. Antibacterial and antibiofilm activity of rAzurin with different concentrations were determined by micro-broth dilution and crystal violet methods, respectively. The effect of rAzurin on bacterial species was statistically analyzed by t- test and spearman correla- tion. Results: The identity of purified protein was confirmed by blotting and distinguished as a 14 kDa band on 15% SDS-PAGE. The IC50 of rAzurin on Peripheral Blood Mononuclear Cell (PBMC) was determined as 377.91±0.5 µg/mL in 24 h. Vibrio cholerae and Campilobacter jejuni displayed the most sensitivity to rAzurin (27.5 and 55 μg/mL, respectively) and the highest resistance (220 μg/mL) was displayed by P. aeruginosa and E. coli. The MIC for other species was 110 μg/mL. The Minimum Biofilm Inhibition Concentration (MBIC) was determined as 220 μg/mL for Salmonella enterica and V. cholerae, 300 μg/mL for Shigella sonnei, Shigella flexneri and P. aeruginosa and 440 μg/mL for the other species. The antimicrobial effect of rAzurin on bacterial species were significant (p value |
| Databáze: | OpenAIRE |
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