Uncovering Endolysins against Methicillin-Resistant Staphylococcus aureus Using a Microbial Single-Cell Genome...

Area of Science:

• Microbiology
• Genomics
• Biotechnology

Background:

• Endolysins, bacteriophage-derived peptidoglycan hydrolases, are promising antibiotic alternatives.
• Developing diverse and active endolysin libraries is crucial for their therapeutic application.

Purpose of the Study:

• To establish microbial single-cell genome sequencing as a method for discovering novel endolysins.
• To identify and characterize new endolysins with antimicrobial activity against Staphylococcus species.

Main Methods:

• Utilized microbial single-cell genome sequencing to identify endolysin genes in Staphylococcus prophages from human skin microbiome samples.
• Constructed chimeric endolysin libraries by domain shuffling.
• Performed high-throughput screening of endolysins against Staphylococcus aureus.

Main Results:

• Successfully recovered previously unknown endolysin genes using single-cell sequencing.
• Identified a lead endolysin, bbst1027, with rapid bactericidal activity against Staphylococcus aureus.
• bbst1027 demonstrated no detectable resistance development and showed in vivo efficacy.

Conclusions:

• Microbial single-cell genome sequencing is an effective approach for discovering novel bacteriophage endolysins.
• The developed endolysin discovery pipeline can be applied to various bacterial targets for novel antimicrobial agent development.