LACTONASE MEDIATED QUORUM QUENCHING OF PSEUDOMONAS AERUGINOSA VIRULENCE
Апстракт
Solving the problem of the antimicrobial resistance crisis is one
of the primary challenges currently confronting the healthcare
system. One of the most promising new strategies to combat
antimicrobial resistance is the antivirulence therapy, based on
silencing bacterial cell-to-cell communication (quorum quenching -
QQ). QQ enzymes lactonases represent a diverse group of enzymes
capable of inactivating signaling molecules of bacterial
communication – N-acyl homoserine lactones (AHLs), resulting in
alterations ofbacterial virulence. The numerous virulence factors and
resistance to most conventional antibiotics have led to Pseudomonas
aeruginosa being listed as one of the top-priority pathogens on the
ESCAPE pathogen list, highlighting the urgent need for the development of new therapies to combat this pathogen. P.
aeruginosauses cell-to-cell communication known as quorum sensing
(QS) that allows bacteria to monitor their own population density via
signal molecules and sub...sequently control bacterial pathogenesis.
Our hypothesis was that bacterial pathogens which share the same
ecological niche with P. aeruginosa during infection have developed a
system to disrupt its QS system, in order to survive in polymicrobial
communities alongside this successful pathogen. In our research we
identified QQ enzymes lactonases originating from two Gramnegative
bacterial pathogens Burkholderiacepacia and Stenotrophomonas
maltophilia. The genes encoding for the enzymes were cloned and
expressed in pQE30 expression vector. B. cepacia BCC4135 synthesizes
two lactonases YtnP and Y2-aiiA, that have the different cellular
localization, but also different substrate specificity, which could imply
the difference in their biological roles. S. maltophilia 6960 YtnP
lactonase has several advantageous biotechnological properties, such
as high thermostability, activity in a wide pH range, and no cytotoxic microscopy analysis showed a strong effect of analyzed
lactonases on preventing biofilm formationand initiating the
decomposition of the preformed biofilm of P. aeruginosa MMA83.
Functional assays showed that lactonases have the ability to
significantly reduce extracellular virulence factors production –
elastase, pyocyanin and rhamnolipid. Additionally, the results
obtained by real-time quantitative PCR showed that analyzed
recombinant enzymes significantly downregulated all three analyzed
P. aeruginosa QS networks at the transcriptional level. Finally, S.
maltophilia 6960 YtnP lactonase significantly prolonged survival of
Caenorhabditis elegans by reducing virulence of P. aeruginosa using fastkilling
liquid assay.
The described properties make B. cepacia and S. maltophilia lactonases
the promising therapeutic candidates for the development of nextgeneration
antivirulence agents.
Кључне речи:
quorum sensing / quorum quenching / lactonase / Pseudomonas aeruginosa / biofilm / virulence / antivirulence therapyИзвор:
3nd International conference Antimicrobial Resistance – current state and perspectives, 2024, 112-114Издавач:
- Novi Sad : Faculty of Agriculture
Финансирање / пројекти:
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200042 (Универзитет у Београду, Институт за молекуларну генетику и генетичко инжењерство) (RS-MESTD-inst-2020-200042)
Напомена:
- Book of abstracts and conference proceedings / 3rd International Conference Antimicrobial Resistance - Current State and Perspectives, 16-18. May 2024, Novi Sad, Serbia.
Колекције
Институција/група
Institut za molekularnu genetiku i genetičko inženjerstvoTY - CONF AU - Malešević, Milka AU - Ćurčić, Jovana AU - Jovčić, Branko PY - 2024 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/2397 AB - Solving the problem of the antimicrobial resistance crisis is one of the primary challenges currently confronting the healthcare system. One of the most promising new strategies to combat antimicrobial resistance is the antivirulence therapy, based on silencing bacterial cell-to-cell communication (quorum quenching - QQ). QQ enzymes lactonases represent a diverse group of enzymes capable of inactivating signaling molecules of bacterial communication – N-acyl homoserine lactones (AHLs), resulting in alterations ofbacterial virulence. The numerous virulence factors and resistance to most conventional antibiotics have led to Pseudomonas aeruginosa being listed as one of the top-priority pathogens on the ESCAPE pathogen list, highlighting the urgent need for the development of new therapies to combat this pathogen. P. aeruginosauses cell-to-cell communication known as quorum sensing (QS) that allows bacteria to monitor their own population density via signal molecules and subsequently control bacterial pathogenesis. Our hypothesis was that bacterial pathogens which share the same ecological niche with P. aeruginosa during infection have developed a system to disrupt its QS system, in order to survive in polymicrobial communities alongside this successful pathogen. In our research we identified QQ enzymes lactonases originating from two Gramnegative bacterial pathogens Burkholderiacepacia and Stenotrophomonas maltophilia. The genes encoding for the enzymes were cloned and expressed in pQE30 expression vector. B. cepacia BCC4135 synthesizes two lactonases YtnP and Y2-aiiA, that have the different cellular localization, but also different substrate specificity, which could imply the difference in their biological roles. S. maltophilia 6960 YtnP lactonase has several advantageous biotechnological properties, such as high thermostability, activity in a wide pH range, and no cytotoxic microscopy analysis showed a strong effect of analyzed lactonases on preventing biofilm formationand initiating the decomposition of the preformed biofilm of P. aeruginosa MMA83. Functional assays showed that lactonases have the ability to significantly reduce extracellular virulence factors production – elastase, pyocyanin and rhamnolipid. Additionally, the results obtained by real-time quantitative PCR showed that analyzed recombinant enzymes significantly downregulated all three analyzed P. aeruginosa QS networks at the transcriptional level. Finally, S. maltophilia 6960 YtnP lactonase significantly prolonged survival of Caenorhabditis elegans by reducing virulence of P. aeruginosa using fastkilling liquid assay. The described properties make B. cepacia and S. maltophilia lactonases the promising therapeutic candidates for the development of nextgeneration antivirulence agents. PB - Novi Sad : Faculty of Agriculture C3 - 3nd International conference Antimicrobial Resistance – current state and perspectives T1 - LACTONASE MEDIATED QUORUM QUENCHING OF PSEUDOMONAS AERUGINOSA VIRULENCE EP - 114 SP - 112 UR - https://hdl.handle.net/21.15107/rcub_imagine_2397 ER -
@conference{ author = "Malešević, Milka and Ćurčić, Jovana and Jovčić, Branko", year = "2024", abstract = "Solving the problem of the antimicrobial resistance crisis is one of the primary challenges currently confronting the healthcare system. One of the most promising new strategies to combat antimicrobial resistance is the antivirulence therapy, based on silencing bacterial cell-to-cell communication (quorum quenching - QQ). QQ enzymes lactonases represent a diverse group of enzymes capable of inactivating signaling molecules of bacterial communication – N-acyl homoserine lactones (AHLs), resulting in alterations ofbacterial virulence. The numerous virulence factors and resistance to most conventional antibiotics have led to Pseudomonas aeruginosa being listed as one of the top-priority pathogens on the ESCAPE pathogen list, highlighting the urgent need for the development of new therapies to combat this pathogen. P. aeruginosauses cell-to-cell communication known as quorum sensing (QS) that allows bacteria to monitor their own population density via signal molecules and subsequently control bacterial pathogenesis. Our hypothesis was that bacterial pathogens which share the same ecological niche with P. aeruginosa during infection have developed a system to disrupt its QS system, in order to survive in polymicrobial communities alongside this successful pathogen. In our research we identified QQ enzymes lactonases originating from two Gramnegative bacterial pathogens Burkholderiacepacia and Stenotrophomonas maltophilia. The genes encoding for the enzymes were cloned and expressed in pQE30 expression vector. B. cepacia BCC4135 synthesizes two lactonases YtnP and Y2-aiiA, that have the different cellular localization, but also different substrate specificity, which could imply the difference in their biological roles. S. maltophilia 6960 YtnP lactonase has several advantageous biotechnological properties, such as high thermostability, activity in a wide pH range, and no cytotoxic microscopy analysis showed a strong effect of analyzed lactonases on preventing biofilm formationand initiating the decomposition of the preformed biofilm of P. aeruginosa MMA83. Functional assays showed that lactonases have the ability to significantly reduce extracellular virulence factors production – elastase, pyocyanin and rhamnolipid. Additionally, the results obtained by real-time quantitative PCR showed that analyzed recombinant enzymes significantly downregulated all three analyzed P. aeruginosa QS networks at the transcriptional level. Finally, S. maltophilia 6960 YtnP lactonase significantly prolonged survival of Caenorhabditis elegans by reducing virulence of P. aeruginosa using fastkilling liquid assay. The described properties make B. cepacia and S. maltophilia lactonases the promising therapeutic candidates for the development of nextgeneration antivirulence agents.", publisher = "Novi Sad : Faculty of Agriculture", journal = "3nd International conference Antimicrobial Resistance – current state and perspectives", title = "LACTONASE MEDIATED QUORUM QUENCHING OF PSEUDOMONAS AERUGINOSA VIRULENCE", pages = "114-112", url = "https://hdl.handle.net/21.15107/rcub_imagine_2397" }
Malešević, M., Ćurčić, J.,& Jovčić, B.. (2024). LACTONASE MEDIATED QUORUM QUENCHING OF PSEUDOMONAS AERUGINOSA VIRULENCE. in 3nd International conference Antimicrobial Resistance – current state and perspectives Novi Sad : Faculty of Agriculture., 112-114. https://hdl.handle.net/21.15107/rcub_imagine_2397
Malešević M, Ćurčić J, Jovčić B. LACTONASE MEDIATED QUORUM QUENCHING OF PSEUDOMONAS AERUGINOSA VIRULENCE. in 3nd International conference Antimicrobial Resistance – current state and perspectives. 2024;:112-114. https://hdl.handle.net/21.15107/rcub_imagine_2397 .
Malešević, Milka, Ćurčić, Jovana, Jovčić, Branko, "LACTONASE MEDIATED QUORUM QUENCHING OF PSEUDOMONAS AERUGINOSA VIRULENCE" in 3nd International conference Antimicrobial Resistance – current state and perspectives (2024):112-114, https://hdl.handle.net/21.15107/rcub_imagine_2397 .