Nisin is widely used in the food industry as a natural preservativedue to its broad antibacterial activity against Gram-positive spoilage and pathogenic bacteria. Using Lactococcus lactis subsp. lactis TCCC12001 as the fermentation strain, culture medium composition was optimized by response surface methodology, which contained 20g/L sucrose, 10g/L yeast, 9g/L peptone, 5g/L beef extract, 10g/L K2HPO4·12H2O, 2g/L NaCl and 0.2g/L MgSO4·7H2O. At optimal fermentation condition (initial pH 7.0, fermentation temperature 30℃, seed age 10h and inoculum density 3%). When cultured in the optimum medium, the nisin yield could reach up to 1436IU/ml, which was 69% higher than that without optimization.