The effects of partial replacement of whole milk powder by wine lees on the characteristics of yogurt by the fermentation of Lactobacillus bulgaricus and Streptococcus thermophiles were evaluated in this study. Single-factor experiment of process conditions, including additive amount of wine lees, additive amount of agar, fermentation time and strains complex ratio were tested to improve the yogurt quality. These major parameters were further optimized by orthogonal experiment. The optimum process conditions were: wine lees amount of 40%, agar of 0.3%, fermentation time of 8 h, and Lactobacillus bulgaricus and Streptococcus thermophiles ratio of 1:1. Under these conditions, the hardness of yogurt was 45.9 g, which is well in close agreement with regular set-style yogurt. This yogurt has the unique fermented flavor of wine lees, and its physicochemical properties and sensory index was similar to regular set-style yogurt.