Anti-virulent Disruption of Pathogenic Biofilms using Engineered Quorum-quenching Lactonases

Tay, Song Buck; Chow, Jeng Yeong; Go, Maybelle Kho; Yew, Wen Shan

doi:10.3791/53243

Cited by 4 publications

(3 citation statements)

References 15 publications

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“…Owing to the continuous emergence of resistant strains to commonly used antibiotics, there is a crucial need for exploring new therapeutic approaches. Lactonases are among the QQ enzymes that had received much more attention in the last few years as promising alternatives targeting AHL molecules of Gram negative pathogens without affecting cell growth or imposing selective pressure on microorganisms 48. They exert their effect by hydrolyzing the ester bond of AHLs, and give the corresponding AHL molecule,19,49 thus preventing the interaction between AHL and its cognate receptor protein and the subsequent expression of virulence phenotypes in pathogenic bacteria.…”

Section: Resultsmentioning

confidence: 99%

Quorum quenching activity of Bacillus cereus isolate 30b confers antipathogenic effects in Pseudomonas aeruginosa

Raafat¹,

Ali-Tammam²,

Ali³

2019

IDR

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Background: Quorum quenching, the interference of a Quorum sensing (QS) system that contributes to the pathogenesis through triggering the production of various virulence determinants, is among the newly suggested antivirulence strategies. Purpose: This study aimed at screening of N-Acyl homoserine lactonase activity from local bacterial isolate, and investigating its effect on Pseudomonas aeruginosa ( P. aeruginosa ) virulence and biofilm formation. Materials and methods: Soil bacteria were screened for aiiA gene coding for lactonase enzyme by Polymerase Chain reaction and sequencing of aiiA gene homologs. Lactonase activity and spectrum were assessed in the cell-free lysate by well diffusion assay using Agrobacterium tumafaciens KYC55. A bacterial isolate showing the highest N-acyl-homoserine lactones degradation percentage was identified by gene amplification and sequencing of the 16S rRNA gene and its aiiA gene homolog. High performance liquid chromatography was used to confirm N-acyl-homoserine lactone degradation. The effect of cell-free lysate on the biofilm formation ability and cytotoxicity of P. aeruginosa PAO1 and P. aeruginosa clinical isolates from different clinical sources were assessed by static microtiter plate and viability assay, respectively Results: Lactonase gene and activity were identified in three Bacillus spp. isolates. They showed broad catalytic activities against tested N-acyl-homoserine lactones. However, The lactonase activity in the cell- free lysate of isolate 30b showed the highest significant degradation percentage on all tested signals; N-butanoyl-L-homoserine lactone (71%), N-hexanoyl- l -homoserine lactone (100%), N-decanoyl-homoserine lactone (100%), N-(3-oxohexanoyl)-L-homoserine lactone (37.5%), N-(oxodecanoyl)-L-homoserine lactone (100%), and N-(3-oxododecanoyl)-L-homoserine lactone (100%). Alignment of the amino acid sequences of AiiA protein of isolate 30b showed 96% identity with Bacillus cereus ( B. cereus ) homologous lactonases in the GenBank database, and the isolate was designated as B. cereus isolate 30b. Cell-free lysate of B. cereus isolate 30b reduced biofilm formation significantly in 93% of P. aeruginosa isolates. The highest mean percentage of reduction in the biofilm was 86%. Moreover, the viability percentage of human lung carcinoma A549 cells infected by P. aeruginosa and treated with cell-free lysate of B. cereus isolate 30b increased up to 15%. Conclusion: The results of this study highlight the potential of lactonases as a promising strategy to combat ...

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Section: Resultsmentioning

confidence: 99%

Quorum quenching activity of Bacillus cereus isolate 30b confers antipathogenic effects in Pseudomonas aeruginosa

Raafat¹,

Ali-Tammam²,

Ali³

2019

IDR

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“…Fifty hours after infection, 75% mortality was observed in untreated rats, whereas mortality was reduced to 20% in treated animals (90). QQ effects were also obtained in A. baumannii, with another engineered enzyme, GKL E101G/ R230C, able to reduce biomass, thickness, and surface of biofilm formed by this pathogen (91,92).…”

Section: Human Healthmentioning

confidence: 99%

Engineering acyl-homoserine lactone-interfering enzymes toward bacterial control

Billot

Plener

Jacquet

et al. 2020

Journal of Biological Chemistry

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Enzymes able to degrade or modify acyl-homoserine lactones (AHL) have drawn considerable interest for their ability to interfere with the bacterial communication process referred to as Quorum Sensing (QS). Many proteobacteria use AHL to coordinate virulence and biofilm formation in a cell density-dependent manner, thus AHL-interfering enzymes constitute new promising antimicrobial candidates. Among these, lactonases and acylases, have been particularly studied. These enzymes have been isolated from various bacterial, archaeal or eukaryotic organisms and have been evaluated for their ability to control several pathogens. Engineering studies on these enzymes were carried out and successfully modulated their capacity to interact with specific AHL, increase their catalytic activity and stability or enhance their biotechnological potential. In this review, special attention is paid to the screening, engineering and applications of AHL-modifying enzymes. Prospects and future opportunities are also discussed with view to developing potent candidates for bacterial control.

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“…New biofilm-control methods, such as inhibition of quorum sensing (QS), enzymatic disruption, bactericidal coating, nanotechnology, and bioelectric approaches, have successfully been studied in an effort to find effective alternatives for the prevention and growth control of biofilms [ 16 , 17 , 18 , 19 , 20 ]. To date, several approaches using different alloys and antimicrobial coatings have been developed with the objective of controlling bacterial adhesion and viability on surfaces.…”

Section: Introductionmentioning

confidence: 99%

Functional Antimicrobial Surface Coatings Deposited onto Nanostructured 316L Food-Grade Stainless Steel

et al. 2021

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In our study, we demonstrated the performance of antimicrobial coatings on properly functionalized and nanostructured 316L food-grade stainless steel pipelines. For the fabrication of these functional coatings, we employed facile and low-cost electrochemical techniques and surface modification processes. The development of a nanoporous structure on the 316L stainless steel surface was performed by following an electropolishing process in an electrolytic bath, at a constant anodic voltage of 40 V for 10 min, while the temperature was maintained between 0 and 10 °C. Subsequently, we incorporated on this nanostructure additional coatings with antimicrobial and bactericide properties, such as Ag nanoparticles, Ag films, or TiO2 thin layers. These functional coatings were grown on the nanostructured substrate by following electroless process, electrochemical deposition, and atomic layer deposition (ALD) techniques. Then, we analyzed the antimicrobial efficiency of these functionalized materials against different biofilms types (Candida parapsilosis, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus epidermidis). The results of the present study demonstrate that the nanostructuring and surface functionalization processes constitute a promising route to fabricate novel functional materials exhibiting highly efficient antimicrobial features. In fact, we have shown that our use of an appropriated association of TiO2 layer and Ag nanoparticle coatings over the nanostructured 316L stainless steel exhibited an excellent antimicrobial behavior for all biofilms examined.

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Anti-virulent Disruption of Pathogenic Biofilms using Engineered Quorum-quenching Lactonases

Cited by 4 publications

References 15 publications

<p>Quorum quenching activity of <em>Bacillus cereus</em> isolate 30b confers antipathogenic effects in <em>Pseudomonas aeruginosa</em></p>

<p>Quorum quenching activity of <em>Bacillus cereus</em> isolate 30b confers antipathogenic effects in <em>Pseudomonas aeruginosa</em></p>

Engineering acyl-homoserine lactone-interfering enzymes toward bacterial control

Functional Antimicrobial Surface Coatings Deposited onto Nanostructured 316L Food-Grade Stainless Steel

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