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Updated: Nov 10, 2025
Bacteriophage Effectiveness for Biocontrol of Foodborne Pathogens Evaluated via High-Throughput Settings
Published on: August 19, 2021
Comparison of Enterococcus faecalis Biofilm Removal Efficiency among Bacteriophage PBEF129, Its Endolysin, and
Hyun Keun Oh1, Yoon Jung Hwang1, Hye Won Hong2
1Department of Bioscience and Biotechnology, Hankuk University of Foreign Studies, Gyung-Gi Do 17035, Korea.
Abstract:
Enterococcus faecalis is a Gram-positive pathogen which colonizes human intestinal surfaces, forming biofilms, and demonstrates a high resistance to many antibiotics. Especially, antibiotics are less effective for eradicating biofilms and better alternatives are needed. In this study, we have isolated and characterized a bacteriophage, PBEF129, infecting E. faecalis. PBEF129 infected a variety of strains of E. faecalis, including those exhibiting antibiotic resistance. Its genome is a linear double-stranded DNA, 144,230 base pairs in length. Its GC content is 35.9%. The closest genomic DNA sequence was found in Enterococcus phage vB_EfaM_Ef2.3, with a sequence identity of 99.06% over 95% query coverage. Furthermore, 75 open reading frames (ORFs) were functionally annotated and five tRNA-encoding genes were found. ORF 6 was annotated as a phage endolysin having an L-acetylmuramoyl-l-alanine amidase activity. We purified the enzyme as a recombinant protein and confirmed its enzymatic activity. The endolysin's host range was observed to be wider than its parent phage PBEF129. When applied to bacterial biofilm on the surface of in vitro cultured human intestinal cells, it demonstrated a removal efficacy of the same degree as cefotaxime, but much lower than its parent bacteriophage.
Insights
A novel bacteriophage, PBEF129, and its endolysin were identified to combat antibiotic-resistant Enterococcus faecalis biofilms. The endolysin showed promising biofilm eradication capabilities, offering a potential alternative to traditional antibiotics.
Area of Science:
- Microbiology
- Bacteriophage Therapy
- Biofilm Research
Background:
- Enterococcus faecalis is a resilient Gram-positive pathogen causing infections and forming antibiotic-resistant biofilms.
- Conventional antibiotics show limited efficacy against E. faecalis biofilms, necessitating alternative therapeutic strategies.
Purpose of the Study:
- To isolate and characterize a bacteriophage targeting E. faecalis.
- To investigate the potential of bacteriophage-derived endolysins as antimicrobial agents against E. faecalis biofilms.
Main Methods:
- Isolation and characterization of bacteriophage PBEF129 infecting E. faecalis.
- Genomic analysis of PBEF129, including ORF annotation.
- Purification and enzymatic activity confirmation of endolysin (ORF 6).
- Assessment of endolysin's host range and biofilm removal efficacy in vitro.
Main Results:
- Bacteriophage PBEF129 was isolated, infecting various E. faecalis strains, including antibiotic-resistant ones.
- PBEF129 possesses a 144,230 bp dsDNA genome with 75 ORFs; endolysin ORF 6 exhibits L-acetylmuramoyl-l-alanine amidase activity.
- The purified endolysin demonstrated a broader host range than the parent phage and comparable biofilm removal efficacy to cefotaxime.
Conclusions:
- Bacteriophage PBEF129 and its endolysin represent promising candidates for combating E. faecalis infections, particularly antibiotic-resistant biofilms.
- Endolysins offer a potential alternative or adjunct therapy for biofilm-associated infections, warranting further investigation.
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