Academic Journal
Molecular characterization of the PhiKo endolysin from Thermus thermophilus HB27 bacteriophage phiKo and its cryptic lytic peptide RAP-29
| العنوان: | Molecular characterization of the PhiKo endolysin from Thermus thermophilus HB27 bacteriophage phiKo and its cryptic lytic peptide RAP-29 |
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| المؤلفون: | Szadkowska, Monika, Kocot, Aleksandra Maria, Sowik, Daria, Wyrzykowski, Dariusz, Jankowska, Elzbieta, Kozlowski, Lukasz Pawel, Makowska, Joanna, Plotka, Magdalena |
| المصدر: | Frontiers in Microbiology ; volume 14 ; ISSN 1664-302X |
| بيانات النشر: | Frontiers Media SA |
| سنة النشر: | 2024 |
| المجموعة: | Frontiers (Publisher - via CrossRef) |
| الوصف: | Introduction In the era of increasing bacterial resistance to antibiotics, new bactericidal substances are sought, and lysins derived from extremophilic organisms have the undoubted advantage of being stable under harsh environmental conditions. The PhiKo endolysin is derived from the phiKo bacteriophage infecting Gram-negative extremophilic bacterium Thermus thermophilus HB27. This enzyme shows similarity to two previously investigated thermostable type-2 amidases, the Ts2631 and Ph2119 from Thermus scotoductus bacteriophages, that revealed high lytic activity not only against thermophiles but also against Gram-negative mesophilic bacteria. Therefore, antibacterial potential of the PhiKo endolysin was investigated in the study presented here. Methods Enzyme activity was assessed using turbidity reduction assays (TRAs) and antibacterial tests. Differential scanning calorimetry was applied to evaluate protein stability. The Collection of Anti-Microbial Peptides (CAMP) and Antimicrobial Peptide Calculator and Predictor (APD3) were used to predict regions with antimicrobial potential in the PhiKo primary sequence. The minimum inhibitory concentration (MIC) of the RAP-29 synthetic peptide was determined against Gram-positive and Gram-negative selected strains, and mechanism of action was investigated with use of membrane potential sensitive fluorescent dye 3,3′-Dipropylthiacarbocyanine iodide (DiSC 3 (5)). Results and discussion The PhiKo endolysin is highly thermostable with melting temperature of 91.70°C. However, despite its lytic effect against such extremophiles as: T. thermophilus , Thermus flavus , Thermus parvatiensis , Thermus scotoductus , and Deinococcus radiodurans , PhiKo showed moderate antibacterial activity against mesophiles. Consequently, its protein sequence was searched for regions with potential antibacterial activity. A highly positively charged region was identified and synthetized (PhiKo 105-133 ). The novel RAP-29 peptide lysed mesophilic strains of staphylococci and Gram-negative bacteria, ... |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | unknown |
| DOI: | 10.3389/fmicb.2023.1303794 |
| DOI: | 10.3389/fmicb.2023.1303794/full |
| الاتاحة: | http://dx.doi.org/10.3389/fmicb.2023.1303794 https://www.frontiersin.org/articles/10.3389/fmicb.2023.1303794/full |
| Rights: | https://creativecommons.org/licenses/by/4.0/ |
| رقم الانضمام: | edsbas.3D6B9880 |
| قاعدة البيانات: | BASE |
| DOI: | 10.3389/fmicb.2023.1303794 |
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