Abstract: Copper (Cu) is widely used in household appliances, medical treatment, catering and other industries worldwide. Besides excellent electrical and thermal conductivity, Cu is well recognized for its outstanding antimicrobial property. Among antimicrobial metals, Cu features easy availability and low cost. Accordingly, copper-containing steels can achieve superior antimicrobial performance after antimicrobial heat treatment. Taking the surface antimicrobial property difference between low-carbon copper-containing steel and ordinary low-carbon steel as the research object, the effects of annealing temperature and Cu content on the antimicrobial performance of low-carbon copper-containing steel were investigated by optical microscopy (OM), ion dissolution test and antimicrobial experiment. The results show that within a certain range, higher Cu content leads to more precipitated Cu particles, higher dissolved Cu ion concentration and better antimicrobial effect. After co-cultivation with Escherichia coli for 24 h, the low-carbon copper-containing steel achieves a 100% sterilization rate against Escherichia coli, which presents favorable application potential in food and medical material fields.