Discovery of Bactericidal Proteins from Staphylococcus Phage Stab21 Using a High-Throughput Screening Method

Area of Science:

• Microbiology
• Genomics
• Biotechnology

Background:

• Antimicrobial resistance necessitates novel antibiotic discovery.
• Bacteriophages represent a promising, yet underexplored, source of antimicrobials.
• Hypothetical Proteins of Unknown Function (HPUFs) in phages are largely uncharacterized for therapeutic potential.

Purpose of the Study:

• To investigate the bactericidal activity of HPUFs from Staphylococcus phage Stab21 against E. coli.
• To identify potential shared molecular targets between E. coli and Staphylococcus aureus.
• To evaluate the utility of Next Generation Sequencing (NGS) for HPUF screening.

Main Methods:

• Screening of 96 HPUFs from Stab21 phage against E. coli.
• Utilized a Next Generation Sequencing (NGS)-based approach for high-throughput analysis.
• Bioinformatic analysis to identify homologous sequences, including homing endonucleases.

Main Results:

• Five out of 96 HPUFs exhibited cross-species toxicity against E. coli.
• Identified one toxic antibacterial HPUF (toxHPUF) homologous to a homing endonuclease.
• Confirmed the effectiveness of NGS in rapidly screening HPUFs for antimicrobial activity.

Conclusions:

• Staphylococcus phage HPUFs can possess broad-spectrum bactericidal activity.
• The identified toxHPUF suggests potential conserved targets relevant to combating Gram-negative bacteria.
• This study validates NGS as an efficient tool for discovering novel phage-derived antimicrobials and advancing phage biology research.