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Bacteriophage P49 is a newly identified, broad-spectrum lytic bacteriophage belonging to the genus *Epseptimavirus* (family *Demerecviridae*). It was isolated from wastewater at West China Hospital, Sichuan University, using phage-resistant mutants of carbapenem-resistant *Klebsiella pneumoniae* (CRKP) as host strains. P49 exhibits a rare cross-genus bactericidal activity, capable of lysing not only phage-resistant CRKP mutants but also *Escherichia coli*, *Salmonella enterica*, *Kluyvera tianfuensis*, and *Enterobacter ludwigii*. It targets and binds to the bacterial outer membrane vitamin B12 transport protein BtuB as its receptor. Due to its stability, rapid adsorption, high burst size, and lack of antimicrobial resistance or virulence genes, bacteriophage P49 is a promising candidate for phage therapy, particularly in combination cocktails to overcome bacterial resistance.
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