Using Escherichia coli O157:H7 as the host bacterium, phages were isolated. Their biological characteristics and genomic features were determined, and their antibacterial effects in food were evaluated. The double-layer plate method was used to separate the bacteriophages. The morphology was observed under a transmission electron microscope. The heat resistance and pH stability were determined. The optimal infection titer was measured and a one-step growth curve was plotted. The genome was sequenced and analyzed. The 24-hour antibacterial effect in different food matrices was evaluated. The bacteriophage isolated from sewage, named ESP02, has a head size of approximately 54 nm × 60 nm and a tail size of about 72 nm. It belongs to the phycocyanophage order and the myovirus family. It has good tolerance at temperatures ranging from 4 ℃~ 60 ℃ and pH levels from 3 ~ 11. The optimal infection multiplicity (MOI) is 0.1, the latent period is 20 minutes, the outbreak period is 20 minutes to 70 minutes, and the outbreak quantity is 88.75 PFU/cell. The complete genome is 48.3 kb in length, containing 76 coding sequences (CDS), with a GC content of 53.27%, and does not contain resistance genes or virulence genes. At 4 ℃ and 25 ℃ conditions, the inhibition rate of the infection multiplicity (MOI = 1000 ~ 10000) for the host bacteria after 24 hours can reach over 99%. The research results show that the virulent phage ESP02 possesses excellent biological properties and safety, as well as effective antibacterial potential against various food matrices under both normal temperature and refrigerated conditions. This also lays the foundation for the development of phage preparations targeting Escherichia coli O157:H7 and their application in food biosecurity control.