TY  - JOUR
AU  - Ćurčić, Jovana
AU  - Dinić, Miroslav
AU  - Novović, Katarina
AU  - Vasiljević, Zorica
AU  - Kojić, Milan
AU  - Jovčić, Branko
AU  - Malešević, Milka
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/864
AB  - Infections caused by multidrug-resistant pathogens are one of the biggest challenges facing the healthcare system today. Quorum quenching (QQ) enzymes have the potential to be used as innovative enzyme-based antivirulence therapeutics to combat infections caused by multidrug-resistant pathogens. The main objective of this research was to describe the novel YtnP lactonase derived from the clinical isolate Stenotrophomonas maltophilia and to investigate its antivirulence potential against multidrug-resistant Pseudomonas aeruginosa MMA83. YtnP lactonase, the QQ enzyme, belongs to the family of metallo-β-lactamases. The recombinant enzyme has several advantageous biotechnological properties, such as high thermostability, activity in a wide pH range, and no cytotoxic effect. High-performance liquid chromatography analysis revealed the activity of recombinant YtnP lactonase toward a wide range of N-acyl-homoserine lactones (AHLs), quorum sensing signaling molecules, with a higher preference for long-chain AHLs. Recombinant YtnP lactonase was shown to inhibit P. aeruginosa MMA83 biofilm formation, induce biofilm decomposition, and reduce extracellular virulence factors production. Moreover, the lifespan of MMA83-infected Caenorhabditis elegans was prolonged with YtnP lactonase treatment. YtnP lactonase showed synergistic inhibitory activity in combination with gentamicin and acted additively with meropenem against MMA83. The described properties make YtnP lactonase a promising therapeutic candidate for the development of next-generation antivirulence agents.
T2  - International Journal of Biological Macromolecules
T1  - A novel thermostable YtnP lactonase from Stenotrophomonas maltophilia inhibits Pseudomonas aeruginosa virulence in vitro and in vivo
SP  - 130421
DO  - 10.1016/j.ijbiomac.2024.130421
ER  - 

@article{
author = "Ćurčić, Jovana and Dinić, Miroslav and Novović, Katarina and Vasiljević, Zorica and Kojić, Milan and Jovčić, Branko and Malešević, Milka",
year = "2024",
abstract = "Infections caused by multidrug-resistant pathogens are one of the biggest challenges facing the healthcare system today. Quorum quenching (QQ) enzymes have the potential to be used as innovative enzyme-based antivirulence therapeutics to combat infections caused by multidrug-resistant pathogens. The main objective of this research was to describe the novel YtnP lactonase derived from the clinical isolate Stenotrophomonas maltophilia and to investigate its antivirulence potential against multidrug-resistant Pseudomonas aeruginosa MMA83. YtnP lactonase, the QQ enzyme, belongs to the family of metallo-β-lactamases. The recombinant enzyme has several advantageous biotechnological properties, such as high thermostability, activity in a wide pH range, and no cytotoxic effect. High-performance liquid chromatography analysis revealed the activity of recombinant YtnP lactonase toward a wide range of N-acyl-homoserine lactones (AHLs), quorum sensing signaling molecules, with a higher preference for long-chain AHLs. Recombinant YtnP lactonase was shown to inhibit P. aeruginosa MMA83 biofilm formation, induce biofilm decomposition, and reduce extracellular virulence factors production. Moreover, the lifespan of MMA83-infected Caenorhabditis elegans was prolonged with YtnP lactonase treatment. YtnP lactonase showed synergistic inhibitory activity in combination with gentamicin and acted additively with meropenem against MMA83. The described properties make YtnP lactonase a promising therapeutic candidate for the development of next-generation antivirulence agents.",
journal = "International Journal of Biological Macromolecules",
title = "A novel thermostable YtnP lactonase from Stenotrophomonas maltophilia inhibits Pseudomonas aeruginosa virulence in vitro and in vivo",
pages = "130421",
doi = "10.1016/j.ijbiomac.2024.130421"
}

Ćurčić, J., Dinić, M., Novović, K., Vasiljević, Z., Kojić, M., Jovčić, B.,& Malešević, M.. (2024). A novel thermostable YtnP lactonase from Stenotrophomonas maltophilia inhibits Pseudomonas aeruginosa virulence in vitro and in vivo. in International Journal of Biological Macromolecules, 130421.
https://doi.org/10.1016/j.ijbiomac.2024.130421

Ćurčić J, Dinić M, Novović K, Vasiljević Z, Kojić M, Jovčić B, Malešević M. A novel thermostable YtnP lactonase from Stenotrophomonas maltophilia inhibits Pseudomonas aeruginosa virulence in vitro and in vivo. in International Journal of Biological Macromolecules. 2024;:130421.
doi:10.1016/j.ijbiomac.2024.130421 .

Ćurčić, Jovana, Dinić, Miroslav, Novović, Katarina, Vasiljević, Zorica, Kojić, Milan, Jovčić, Branko, Malešević, Milka, "A novel thermostable YtnP lactonase from Stenotrophomonas maltophilia inhibits Pseudomonas aeruginosa virulence in vitro and in vivo" in International Journal of Biological Macromolecules (2024):130421,
https://doi.org/10.1016/j.ijbiomac.2024.130421 . .

TY  - CONF
AU  - Ćurčić, Jovana
AU  - Jakovljević, Stefan
AU  - Novović, Katarina
AU  - Vasiljević, Zorica
AU  - Kojić, Milan
AU  - Jovčić, Branko
AU  - Malešević, Milka
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/803
AB  - Introduction: Quorum quenching (QQ) isthe enzymatic degradation of cell-to-cellsignaling molecules.
In this study, the potential of the novel YtnP lactonase, the quorum quenching enzyme derived from S.
maltophilia, to reduce P. aeruginosa quorum sensing and virulence factor gene expression was investigated.
Methods: MMA83 culture (adjusted to 1.5x105 CFU/ml) was treated with recombinant YtnP lactonase
(final concentration 50 μg/ml) at 37°C for 12 hours under aeration. RNA isolation of the treated and untreated MMA83 culture was performed using the RNeasy Mini Kit (Qiagen, Germany) according to the
protocol. Quantitative reverse transcription-polymerase chain reaction (RT-qPCR), was used to analyze
the effect ofYtnP lactonase on the relative mRNA levels of the LasI/LasR, RhiI/RhiR, and PQS signaling network genes of P. aeruginosa MMA83 and virulence factor genes. The rpsL was used as an endogenous
control to normalize obtained data following the 2-ΔΔCt method.
Results: The QS genes belonging to three QS networks – LasI/LasR, RhiI/RhiR, and PQS of P. aeruginosa
MMA83 treated with YtnP lactonase were significantly downregulated. The RT -qPCR results show that
treatment with YtnP-lactonase decreased the relative mRNA levels of genes involved in the production
of elastase (lasB approximately 2-fold), alginate (algK approximately 2.2-fold), pyocyanin (phzM approximately 3.5-fold), pyoverdin (pvdS approximately 2-fold), and rhamnolipid (rhlC approximately 4-fold).
These results suggest that YtnP lactonase exerts an antivirulence effect at the transcription level.
Conclusion: YtnP lactonase, a quorum quenching (QQ) enzyme, has the potential to be used as an innovative enzyme-based antivirulence therapeutic to combat infections caused by P. aeruginosa.
PB  - Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade
C3  - CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
T1  - A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression
EP  - 121
SP  - 121
UR  - https://hdl.handle.net/21.15107/rcub_intor_803
ER  - 

@conference{
author = "Ćurčić, Jovana and Jakovljević, Stefan and Novović, Katarina and Vasiljević, Zorica and Kojić, Milan and Jovčić, Branko and Malešević, Milka",
year = "2023",
abstract = "Introduction: Quorum quenching (QQ) isthe enzymatic degradation of cell-to-cellsignaling molecules.
In this study, the potential of the novel YtnP lactonase, the quorum quenching enzyme derived from S.
maltophilia, to reduce P. aeruginosa quorum sensing and virulence factor gene expression was investigated.
Methods: MMA83 culture (adjusted to 1.5x105 CFU/ml) was treated with recombinant YtnP lactonase
(final concentration 50 μg/ml) at 37°C for 12 hours under aeration. RNA isolation of the treated and untreated MMA83 culture was performed using the RNeasy Mini Kit (Qiagen, Germany) according to the
protocol. Quantitative reverse transcription-polymerase chain reaction (RT-qPCR), was used to analyze
the effect ofYtnP lactonase on the relative mRNA levels of the LasI/LasR, RhiI/RhiR, and PQS signaling network genes of P. aeruginosa MMA83 and virulence factor genes. The rpsL was used as an endogenous
control to normalize obtained data following the 2-ΔΔCt method.
Results: The QS genes belonging to three QS networks – LasI/LasR, RhiI/RhiR, and PQS of P. aeruginosa
MMA83 treated with YtnP lactonase were significantly downregulated. The RT -qPCR results show that
treatment with YtnP-lactonase decreased the relative mRNA levels of genes involved in the production
of elastase (lasB approximately 2-fold), alginate (algK approximately 2.2-fold), pyocyanin (phzM approximately 3.5-fold), pyoverdin (pvdS approximately 2-fold), and rhamnolipid (rhlC approximately 4-fold).
These results suggest that YtnP lactonase exerts an antivirulence effect at the transcription level.
Conclusion: YtnP lactonase, a quorum quenching (QQ) enzyme, has the potential to be used as an innovative enzyme-based antivirulence therapeutic to combat infections caused by P. aeruginosa.",
publisher = "Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade",
journal = "CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia",
title = "A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression",
pages = "121-121",
url = "https://hdl.handle.net/21.15107/rcub_intor_803"
}

Ćurčić, J., Jakovljević, S., Novović, K., Vasiljević, Z., Kojić, M., Jovčić, B.,& Malešević, M.. (2023). A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 121-121.
https://hdl.handle.net/21.15107/rcub_intor_803

Ćurčić J, Jakovljević S, Novović K, Vasiljević Z, Kojić M, Jovčić B, Malešević M. A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia. 2023;:121-121.
https://hdl.handle.net/21.15107/rcub_intor_803 .

Ćurčić, Jovana, Jakovljević, Stefan, Novović, Katarina, Vasiljević, Zorica, Kojić, Milan, Jovčić, Branko, Malešević, Milka, "A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression" in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia (2023):121-121,
https://hdl.handle.net/21.15107/rcub_intor_803 .