title: Purification and antibiofilm activity of AHL-lactonase from endophytic Enterobacter aerogenes VT66.
abstract: The opportunistic pathogen Pseudomonas aeruginosa uses biofilm lifestyle to resist antibiotic treatment. In our study, endophytic bacterium Enterobacter aerogenes VT66 quenched the N-acyl homoserine lactone (AHL) molecules produced by P. aeruginosa PAO1. The quorum quenching activity was attributed to the presence of AHL-lactonase. The AHL-lactonase was purified using column chromatography and purified AHL-lactonase was applied for the control of biofilm formation in P. aeruginosa PAO1. The results showed that purified AHL-lactonase obtained with a molecular weight about 30kDa was able to inhibit more than 70% of biofilm in P. aeruginosa PAO1 (P<0.001). Antibiofilm activity of AHL-lactonase was correlated well with results from staining technique used to determine inhibition of biomass and viable cell activity. Therefore, results unambiguously confirm that the AHL-lactonase from E. aerogenes VT66 could be used as antibiofilm therapeutics in P. aeruginosa associated biomedical applications.
authors.affiliation: Department of Studies in Microbiology, University of Mysore, Manasagangotri, Mysore 570006, India.
authors.forename: P S
authors.initials: PS
authors.lastname: Rajesh
authors.affiliation: Department of Studies in Microbiology, University of Mysore, Manasagangotri, Mysore 570006, India. Electronic address: raivittal@gmail.com.
authors.forename: V Ravishankar
authors.initials: VR
authors.lastname: Rai
mesh_terms: D018441:Biofilms / Q000187:drug effects*; D002265:Carboxylic Ester Hydrolases / Q000302:isolation & purification* / Q000494:pharmacology*; D002413:Cations, Divalent / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D060026:Endophytes / Q000201:enzymology*; D021902:Enterobacter aerogenes / Q000201:enzymology*; D004795:Enzyme Stability / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007211:Indoles / Q000378:metabolism; D008670:Metals / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D059748:Proteolysis / Q000187:drug effects; D011550:Pseudomonas aeruginosa / Q000502:physiology; D013194:Staining and Labeling; D013379:Substrate Specificity / Q000187:drug effects; D013696:Temperature; D013997:Time Factors
chemical_list: D002413:Cations, Divalent; D007211:Indoles; D008670:Metals; C007293:DAPI; D004492:Edetic Acid; D002265:Carboxylic Ester Hydrolases; C432003:N-acyl homoserine lactonase
keywords: AHL-lactonase; Antibiofilm; Quorum quenching