Analysis of antibacterial and antibiofilm activity of purified recombinant Azurin from Pseudomonas aeruginosa

Autor: Hajar Mohammadi-Barzelighi, Bahram Nasr-Esfahani, Bita Bakhshi, Bahram Daraei, Sharareh Moghim, Hossein Fazeli
Jazyk: angličtina
Rok vydání: 2019
Předmět:
Zdroj: Iranian Journal of Microbiology, Vol 11, Iss 2 (2019)
Druh dokumentu: article
ISSN: 2008-3289
2008-4447
DOI: 10.18502/ijm.v11i2.1083
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 confirmed 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 correlation. 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
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