Analysis of antibacterial and antibiofilm activity of purified recombinant Azurin from Pseudomonas aeruginosa
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...
Main Authors: | , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Tehran University of Medical Sciences
2019-05-01
|
Series: | Iranian Journal of Microbiology |
Subjects: | |
Online Access: | https://ijm.tums.ac.ir/index.php/ijm/article/view/2017 |
id |
doaj-d299f4452d034ae790a5a4e729dcef48 |
---|---|
record_format |
Article |
spelling |
doaj-d299f4452d034ae790a5a4e729dcef482020-12-02T06:31:22ZengTehran University of Medical SciencesIranian Journal of Microbiology2008-32892008-44472019-05-0111210.18502/ijm.v11i2.1083Analysis of antibacterial and antibiofilm activity of purified recombinant Azurin from Pseudomonas aeruginosaHajar Mohammadi-Barzelighi0Bahram Nasr-Esfahani1Bita Bakhshi2Bahram Daraei3Sharareh Moghim4Hossein Fazeli5Department of Microbiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, IranDepartment of Microbiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, IranDepartment of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranToxicology and Pharmacology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, IranDepartment of Microbiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, IranDepartment of Microbiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran 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<0.05) and correlation coefficient was negative. Conclusion: The rAzurin appears to be an appropriate choice and a new strategy for prevention of bacterial infection. It inhibits bacterial growth and biofilm formation and candidates as antimicrobial peptides. https://ijm.tums.ac.ir/index.php/ijm/article/view/2017Antibacterial effectAntibiofilm activityRecombinant Azurin |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Hajar Mohammadi-Barzelighi Bahram Nasr-Esfahani Bita Bakhshi Bahram Daraei Sharareh Moghim Hossein Fazeli |
spellingShingle |
Hajar Mohammadi-Barzelighi Bahram Nasr-Esfahani Bita Bakhshi Bahram Daraei Sharareh Moghim Hossein Fazeli Analysis of antibacterial and antibiofilm activity of purified recombinant Azurin from Pseudomonas aeruginosa Iranian Journal of Microbiology Antibacterial effect Antibiofilm activity Recombinant Azurin |
author_facet |
Hajar Mohammadi-Barzelighi Bahram Nasr-Esfahani Bita Bakhshi Bahram Daraei Sharareh Moghim Hossein Fazeli |
author_sort |
Hajar Mohammadi-Barzelighi |
title |
Analysis of antibacterial and antibiofilm activity of purified recombinant Azurin from Pseudomonas aeruginosa |
title_short |
Analysis of antibacterial and antibiofilm activity of purified recombinant Azurin from Pseudomonas aeruginosa |
title_full |
Analysis of antibacterial and antibiofilm activity of purified recombinant Azurin from Pseudomonas aeruginosa |
title_fullStr |
Analysis of antibacterial and antibiofilm activity of purified recombinant Azurin from Pseudomonas aeruginosa |
title_full_unstemmed |
Analysis of antibacterial and antibiofilm activity of purified recombinant Azurin from Pseudomonas aeruginosa |
title_sort |
analysis of antibacterial and antibiofilm activity of purified recombinant azurin from pseudomonas aeruginosa |
publisher |
Tehran University of Medical Sciences |
series |
Iranian Journal of Microbiology |
issn |
2008-3289 2008-4447 |
publishDate |
2019-05-01 |
description |
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<0.05) and correlation coefficient was negative.
Conclusion: The rAzurin appears to be an appropriate choice and a new strategy for prevention of bacterial infection. It inhibits bacterial growth and biofilm formation and candidates as antimicrobial peptides.
|
topic |
Antibacterial effect Antibiofilm activity Recombinant Azurin |
url |
https://ijm.tums.ac.ir/index.php/ijm/article/view/2017 |
work_keys_str_mv |
AT hajarmohammadibarzelighi analysisofantibacterialandantibiofilmactivityofpurifiedrecombinantazurinfrompseudomonasaeruginosa AT bahramnasresfahani analysisofantibacterialandantibiofilmactivityofpurifiedrecombinantazurinfrompseudomonasaeruginosa AT bitabakhshi analysisofantibacterialandantibiofilmactivityofpurifiedrecombinantazurinfrompseudomonasaeruginosa AT bahramdaraei analysisofantibacterialandantibiofilmactivityofpurifiedrecombinantazurinfrompseudomonasaeruginosa AT shararehmoghim analysisofantibacterialandantibiofilmactivityofpurifiedrecombinantazurinfrompseudomonasaeruginosa AT hosseinfazeli analysisofantibacterialandantibiofilmactivityofpurifiedrecombinantazurinfrompseudomonasaeruginosa |
_version_ |
1724408614316146688 |