Escherichia coli is a common foodborne pathogen that can result in life-threatening diseases and represents a serious burden to food safety management. With the advent of the post-antibiotic era, bacteriophages have generated great attention as promising biocontrol agents for their selective action against bacteria and lysis them. Here, we describe the isolation and charcaterization of a new E. coli-infecting phage DSI29, isolated from a freshwater sample in Joypurhat, Bangladesh. DSI29 phage has a wide host range and a lytic performance to E. coli, implying that the phage is applicable for food safety. Subsequent work revealed its stability to different pH and temperatures, rendering it appropriate for a broad range of environments. Because of its pure lytic lifestyle and powerful bacteriolytic activity, the DSI29 has promising potential as a biocontrol agent in food manufacturing systems. These results reveal the potential of phage therapies in controlling E. coli contamination, a sustainable alternative to conventional antibiotics.