The creation of probiotic-containing fermented milk products for use by human is an important research topic and has high potential for development. Also, studies have shown that heat-killed probiotics are more stable and easier to use than live probiotics. However, as of this time, research has not been reported in Korea that has evaluated the product or functionality of fermented milk after the addition of heat-killed probiotics. This study was conducted to verify the physiological activity of heat-killed Enterococcus faecalis EF-2001 after its addition to Lactobacillus-fermented milk. Briefly, NFM normal fermented milk (NFM) was used as the control sample, whereas fermented milk with 100 μg/mL EF-2001 (EFM1) and fermented milk with 500 μg/mL EF-2001 (EFM2) were used as the treated samples. Among the samples, EFM2 had the highest acidity of 1.15, but no other factors significantly differed (p<0.05). Furthermore, EFM2 had the highest Lactobacillus count of 9.22 (p<0.05). ABTS, DPPH and FRAP were measured to determine the antioxidant activity of the samples. With respect to those parameters, EFM2 had the highest antioxidant measurements. Therefore, the study confirmed that the addition of E. faecalis EF-2001 to NFM is suitable with the standard and does not affect the quality of characteristics. In conclusion, the treatment sample had higher antioxidant activity than did NFM; this result may be used as a basic for further research and as a guideline for the manufacturing of heat-killed probiotic-containing NFM.