Deep eutectic solvent enhances antibacterial activity of a modular lytic enzyme against Acinetobacter baumannii
Abstract In this study, we evaluated the combined effect between MLE-15, a modular lytic enzyme composed of four building blocks, and reline, a natural deep eutectic solvent. The bioinformatic analysis allowed us to determine the spatial architecture of MLE-15, whose components were bactericidal pep...
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Nature Portfolio
2025-01-01
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| Online Access: | https://doi.org/10.1038/s41598-024-80440-z |
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| author | Aleksandra Maria Kocot Tomasz Swebocki Karolina Ciemińska Adrianna Łupkowska Małgorzata Kapusta Dennis Grimon Ewa Laskowska Anna-Karina Kaczorowska Tadeusz Kaczorowski Rabah Boukherroub Yves Briers Magdalena Plotka |
| author_facet | Aleksandra Maria Kocot Tomasz Swebocki Karolina Ciemińska Adrianna Łupkowska Małgorzata Kapusta Dennis Grimon Ewa Laskowska Anna-Karina Kaczorowska Tadeusz Kaczorowski Rabah Boukherroub Yves Briers Magdalena Plotka |
| author_sort | Aleksandra Maria Kocot |
| collection | DOAJ |
| description | Abstract In this study, we evaluated the combined effect between MLE-15, a modular lytic enzyme composed of four building blocks, and reline, a natural deep eutectic solvent. The bioinformatic analysis allowed us to determine the spatial architecture of MLE-15, whose components were bactericidal peptide cecropin A connected via a flexible linker to the cell wall binding domain (CBD) of mesophilic 201ϕ2 − 1 endolysin and catalytic domain (EAD) of highly thermostable Ph2119 endolysin. The modular enzyme showed high thermostability with the melting temperature of 93.97 ± 0.38 °C, significantly higher than their natural counterparts derived from mesophilic sources. The minimum inhibitory concentration (MIC) of MLE-15 was 100 µg/mL for a panel of Gram-positive and Gram-negative bacteria, while the MIC of reline ranged from 6.25 to 25% v/v for the same strains. The addition of reline effectively reduced the MIC of MLE-15 from 100 µg/mL to 3.15–50 µg/mL. This combination displayed additive effects for most strains and synergism for extensively antibiotic-resistant Acinetobacter baumannii and Bacillus subtilis. The subsequent evaluation revealed that MLE-15 eliminated planktonic cells of A. baumannii RUH134, but was ineffective against matured biofilms. However, combined with reline, MLE-15 reduced the bacterial load in the matured biofilm by 1.39 log units. Confocal fluorescence microscopy indicated that reline damaged the structure of the biofilm, allowing MLE-15 to penetrate it. Additionally, MLE-15 and its combination with reline eradicated meropenem-persistent cells of A. baumannii RUH134. Effectiveness in lowering the MIC value of MLE-15 as well as protection against antibiotic-tolerant persister cells, indicate that MLE-15 and reline combination is a promising candidate for effective therapies in bacterial infections, which is especially important in the light of the global crisis of antimicrobial resistance. |
| format | Article |
| id | doaj-art-87b855b55c2b4b489ed362ccfe26a024 |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
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| spelling | doaj-art-87b855b55c2b4b489ed362ccfe26a0242025-01-19T12:17:18ZengNature PortfolioScientific Reports2045-23222025-01-0115111810.1038/s41598-024-80440-zDeep eutectic solvent enhances antibacterial activity of a modular lytic enzyme against Acinetobacter baumanniiAleksandra Maria Kocot0Tomasz Swebocki1Karolina Ciemińska2Adrianna Łupkowska3Małgorzata Kapusta4Dennis Grimon5Ewa Laskowska6Anna-Karina Kaczorowska7Tadeusz Kaczorowski8Rabah Boukherroub9Yves Briers10Magdalena Plotka11Laboratory of Extremophiles Biology, Department of Microbiology, Faculty of Biology, University of GdanskUniv. Lille, CNRS, Univ. Polytechnique Hauts-de-France, UMR 8520 IEMN – Institut d’Electronique de Microélectronique et de NanotechnologieLaboratory of Extremophiles Biology, Department of Microbiology, Faculty of Biology, University of GdanskDepartment of General and Medical Biochemistry, Faculty of Biology, University of GdanskBioimaging Laboratory, Faculty of Biology, University of GdanskDepartment of Biotechnology, Faculty of Bioscience Engineering, Ghent UniversityDepartment of General and Medical Biochemistry, Faculty of Biology, University of GdanskCollection of Plasmids and Microorganisms | KPD, Faculty of Biology, University of GdanskLaboratory of Extremophiles Biology, Department of Microbiology, Faculty of Biology, University of GdanskUniv. Lille, CNRS, Univ. Polytechnique Hauts-de-France, UMR 8520 IEMN – Institut d’Electronique de Microélectronique et de NanotechnologieDepartment of Biotechnology, Faculty of Bioscience Engineering, Ghent UniversityLaboratory of Extremophiles Biology, Department of Microbiology, Faculty of Biology, University of GdanskAbstract In this study, we evaluated the combined effect between MLE-15, a modular lytic enzyme composed of four building blocks, and reline, a natural deep eutectic solvent. The bioinformatic analysis allowed us to determine the spatial architecture of MLE-15, whose components were bactericidal peptide cecropin A connected via a flexible linker to the cell wall binding domain (CBD) of mesophilic 201ϕ2 − 1 endolysin and catalytic domain (EAD) of highly thermostable Ph2119 endolysin. The modular enzyme showed high thermostability with the melting temperature of 93.97 ± 0.38 °C, significantly higher than their natural counterparts derived from mesophilic sources. The minimum inhibitory concentration (MIC) of MLE-15 was 100 µg/mL for a panel of Gram-positive and Gram-negative bacteria, while the MIC of reline ranged from 6.25 to 25% v/v for the same strains. The addition of reline effectively reduced the MIC of MLE-15 from 100 µg/mL to 3.15–50 µg/mL. This combination displayed additive effects for most strains and synergism for extensively antibiotic-resistant Acinetobacter baumannii and Bacillus subtilis. The subsequent evaluation revealed that MLE-15 eliminated planktonic cells of A. baumannii RUH134, but was ineffective against matured biofilms. However, combined with reline, MLE-15 reduced the bacterial load in the matured biofilm by 1.39 log units. Confocal fluorescence microscopy indicated that reline damaged the structure of the biofilm, allowing MLE-15 to penetrate it. Additionally, MLE-15 and its combination with reline eradicated meropenem-persistent cells of A. baumannii RUH134. Effectiveness in lowering the MIC value of MLE-15 as well as protection against antibiotic-tolerant persister cells, indicate that MLE-15 and reline combination is a promising candidate for effective therapies in bacterial infections, which is especially important in the light of the global crisis of antimicrobial resistance.https://doi.org/10.1038/s41598-024-80440-zModular lytic enzymeEndolysinDeep eutectic solventSynergismAntibacterial effectBiofilm |
| spellingShingle | Aleksandra Maria Kocot Tomasz Swebocki Karolina Ciemińska Adrianna Łupkowska Małgorzata Kapusta Dennis Grimon Ewa Laskowska Anna-Karina Kaczorowska Tadeusz Kaczorowski Rabah Boukherroub Yves Briers Magdalena Plotka Deep eutectic solvent enhances antibacterial activity of a modular lytic enzyme against Acinetobacter baumannii Scientific Reports Modular lytic enzyme Endolysin Deep eutectic solvent Synergism Antibacterial effect Biofilm |
| title | Deep eutectic solvent enhances antibacterial activity of a modular lytic enzyme against Acinetobacter baumannii |
| title_full | Deep eutectic solvent enhances antibacterial activity of a modular lytic enzyme against Acinetobacter baumannii |
| title_fullStr | Deep eutectic solvent enhances antibacterial activity of a modular lytic enzyme against Acinetobacter baumannii |
| title_full_unstemmed | Deep eutectic solvent enhances antibacterial activity of a modular lytic enzyme against Acinetobacter baumannii |
| title_short | Deep eutectic solvent enhances antibacterial activity of a modular lytic enzyme against Acinetobacter baumannii |
| title_sort | deep eutectic solvent enhances antibacterial activity of a modular lytic enzyme against acinetobacter baumannii |
| topic | Modular lytic enzyme Endolysin Deep eutectic solvent Synergism Antibacterial effect Biofilm |
| url | https://doi.org/10.1038/s41598-024-80440-z |
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