<p>Bacteriophages (phages), as natural predators of bacteria, represent promising alternatives to antibiotics. However, challenges such as narrow host range and insufficient pharmacokinetic and pharmacodynamic (PK/PD) studies restrict phage therapy. In previous work, we isolated <i>Escherichia</i> phage vB_EcoStr-FJ63A, which exhibits a lytic activity against colistin-resistant <i>Escherichia coli</i> and obvious antibacterial activity in vitro. To elucidate its infection mechanism and evaluate its therapeutic potential, this study focused on identifying its host receptors and assessing its in vivo efficacy. By screening resistant mutants and analyzing the function of mutant sites, we hypothesized that outer membrane proteins (OMPs), lipopolysaccharide (LPS), or extracellular polysaccharides likely serve as receptor candidates for phage vB_EcoStr-FJ63A. The role of LPS as the receptor was supported by competitive binding and receptor removal assays, which demonstrated its necessity for phage adsorption. Bioinformatics analysis results provide evidences for long tail fiber adhesin of phage vB_EcoStr-FJ63A reversibly binds to OmpC, while its short tail fibers irreversibly anchor to LPS. Subsequent PK experiment in healthy mice revealed that a single intraperitoneal injection of phage vB_EcoStr-FJ63A (5 × 10⁸ PFU/mL or 5 × 10⁶ PFU/mL) achieved peak blood titers at 2&#xa0;h post-administration, with phage remaining detectable for over 24&#xa0;h (<i>n</i> = 6 per group at each time point). In bacteremic mice infected with colistin-resistant <i>E. coli</i>, both phage doses significantly reduced bacterial loads in the blood within 8&#xa0;h (<i>n</i> = 3 per group at each time point) and improved survival rates (<i>n</i> = 6 per group) compared with the untreated control group. Collectively, our results provide evidence for the molecular basis of phage–host interactions for vB_EcoStr-FJ63A and provides data to advance phage therapy against antimicrobial resistant <i>E. coli</i>.</p>