Novel endolysin of S. sciuri phage S10 and its antimicrobial effect on S. aureus

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F22%3A00129692" target="_blank" >RIV/00216224:14310/22:00129692 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Novel endolysin of S. sciuri phage S10 and its antimicrobial effect on S. aureus

Popis výsledku v původním jazyce

A lot of novel bacteriophages were isolated, characterised and added to the microbial collection of Fagofarma (and MB Pharma) recently. Some of these phages are unique and their genomes encode potentially novel peptidoglycan hydrolases. Such as the phage S. sciuri S10, which was isolated from wastewater and its genome showed low similarity to sequences found in public databases. In its genome, two genes encoding hypothetical endolysin (LysS10) or tail peptidoglycan hydrolase (TPH) were identified. In this study, we focused on examination of potential activity of these two enzymes. Both genes were cloned, the proteins were expressed in E. coli and purified using chromatography. Zymogram was used as the first method for determination of activity on bacterial cell walls isolated from S. sciuri and S. aureus. This test proved activity of endolysin LysS10 on both types of cell walls. Activity of TPH was not proven, therefore, we focused mainly on LysS10 which activity was verified using spot assay and turbidity reduction assay. Moreover, LysS10 activity was compared with the LysK and the LysF1 on different strains and species of Staphylococcus, including MRSA. LysS10 had antimicrobial effect on every tested bacterium, but it was less active compared to LysK and LysF1. Antimicrobial effect of novel endolysin LysS10 was proven by different methods on several S. aureus strains and other staphylococcal species. Although the enzyme is not as efficient as LysK, there is potential for improving its properties by changing domains, protein design of the catalytic domain or increasing the solubility of the protein.

Název v anglickém jazyce

Novel endolysin of S. sciuri phage S10 and its antimicrobial effect on S. aureus

Popis výsledku anglicky

A lot of novel bacteriophages were isolated, characterised and added to the microbial collection of Fagofarma (and MB Pharma) recently. Some of these phages are unique and their genomes encode potentially novel peptidoglycan hydrolases. Such as the phage S. sciuri S10, which was isolated from wastewater and its genome showed low similarity to sequences found in public databases. In its genome, two genes encoding hypothetical endolysin (LysS10) or tail peptidoglycan hydrolase (TPH) were identified. In this study, we focused on examination of potential activity of these two enzymes. Both genes were cloned, the proteins were expressed in E. coli and purified using chromatography. Zymogram was used as the first method for determination of activity on bacterial cell walls isolated from S. sciuri and S. aureus. This test proved activity of endolysin LysS10 on both types of cell walls. Activity of TPH was not proven, therefore, we focused mainly on LysS10 which activity was verified using spot assay and turbidity reduction assay. Moreover, LysS10 activity was compared with the LysK and the LysF1 on different strains and species of Staphylococcus, including MRSA. LysS10 had antimicrobial effect on every tested bacterium, but it was less active compared to LysK and LysF1. Antimicrobial effect of novel endolysin LysS10 was proven by different methods on several S. aureus strains and other staphylococcal species. Although the enzyme is not as efficient as LysK, there is potential for improving its properties by changing domains, protein design of the catalytic domain or increasing the solubility of the protein.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10600 - Biological sciences

Projekt

<a href="/cs/project/NU21J-05-00035" target="_blank" >NU21J-05-00035: Synergie lytických bakteriofágů a antibiotik v léčbě povrchových infekcí způsobených Staphylococcus aureus</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2022

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů