Aeromonas bakteriofago KLEA5 uodegėlės baltymo gp45 tyrimai /

The widespread availability and overuse of antibiotics is becoming a critical problem due to the growing prevalence of bacterial resistance. In addition, bacteria can form biofilms which act as a virulence factor and prevent drugs from reaching the cell surface, further complicating the issue. The use of bacteriophage-encoded depolymerases offers a promising avenue to address this challenge. These depolymerases may have a wide range of applications in various fields, including biotechnology, industry, and medicine. They are able to degrade polymeric substances present on bacterial surfaces and biofilms. However, they remain underutilised. The aim of this study was to examine the bacteriophage vB_AveS_KLEA5 (KLEA5) tail protein gp45, which exhibits depolymerase properties. The gp45 protein of the KLEA5 phage that infects the Aeromonas veronii KR2-5 isolate has been studied. KLEA5_gp45_pET21a and KLEA5_gp45_pCDFDuet-1 gene expression vectors were amplified. Soluble ~70 kDa gp45_N-his and gp45_C-his proteins were obtained by gene expression in E. coli BL21(DE-3) cells. These proteins are stable and remain active even after 4 months of storage at -20 °C and 5 months of storage at 30 °C. The specificity of the recombinant gp45_N-his and gp45_C-his proteins and of bacteriophage KLEA5 for bacterial cells was determined, and it was found that bacteriophage KLEA5 and its recombinant gp45_N-his and gp45_C-his proteins have a narrow spectrum of activity and that they are specific only for cells of the A. veronii KR2-5 isolate. The study of the interaction of the recombinant proteins with bacterial cells showed that the proteins with depolymerase activity potentially are not receptor binding proteins, but likely hydrolyse A. veronii KR2-5 LPS, resulting in cell lysis. The study of the effect of the recombinant proteins on bacterial cell viability showed that recombinant gp45_N-his reduced the cell viability of A. veronii KR2-5 cells under the conditions studied most significantly after 1.5 h (98.06% reduction in viability), whereas gp45_C-his reduced cell viability after 2 h of incubation (96.92% reduction in viability).

Grantor dc:publisher

Institutional Repository of Vilnius University

Year dc:date

2024

Author dc:creator

• Vedegytė, Miglė,

Statement dc:rights

• info:eu-repo/semantics/openAccess

Language dc:language

lit

Repository record dc:identifier

https://repository.vu.lt/VU:ELABAETD210640164&prefLang=en_US

OAI identifier oai:identifier

oai:vu.lt:elaba:210640164