Lactic acid bacteria are probiotics in the intestines and have been widely used as natural antioxidants in the food industry. In this study, Enterococcus italicus FM5 with strong antioxidant ability was isolated from fresh milk. The safety evaluation showed that E. italicus FM5 was sensitive to ampicillin, chloramphenicol, erythromycin, vancomycin, rifampicin, and tetracycline, and was not hemolytic. Meanwhile, the whole genome information and biofunctional attributes of this strain were determined and analyzed. Subsequently, E. italicus FM5 was co-cultured with traditional yogurt starters (Streptococcus thermophilus and Lactobacillus bulgaricus) to make fermented milk. The results showed that the addition of E. italicus FM5 could improve the oxygen free radical scavenging ability of the fermented milk, and the scavenging rates of DPPH, ABTS, OH