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Ciemińska, Karolina, 2024, "Boosting the Antibacterial Activity of Phage-Derived Modular Enzyme MLE-15 with Natural Deep Eutectic Solvent as a Co-Drug Against Multidrug-Resistant Acinetobacter baumannii RUH134", https://doi.org/10.18150/OWFIIK, RepOD, V1
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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 3.15 to 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 entirely prevented the formation of a biofilm and eradicated meropenem-persistent cells of A. baumannii RUH134. Effectiveness in lowering the MIC value of MLE-15 as well as complete protection against biofilm formation and 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.
modular lytic enzyme, endolysin, deep eutectic solvent, synergism, antibacterial effect, biofilm, persisters
CC BY - Creative Commons Attribution 4.0
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