The production of nisin by batch culture of Lactococcus lactis ATCC 11454 in MRS broth (pH 6.5), as treated in 30 assays, that were set up by a fractional factorial design of two levels (2[4–1]), was improved. The minimum and maximum concentrations of sucrose (5.0–12.5 g/L), asparagine (7.5–75 g/L), potassium phosphate (6.0–18.0 g/L), and Tween-80 (1.0–6.6 g/L) were added to MRS broth. The best nisin activities ranged from 1.5×104 to 1.8×104 arbitrary units (AU)/mL for the maximum levels of sucrose, asparagine, and monobasic potassium phosphate, and for the minimum concentration of Tween-80. The best following proportions between nutrients were adopted as optimum for maximum specific nisin productivity of about 6.0 mg/mg of dry cell weight (related to 2.5 mg of pure nisin preparation with a specific activity of 1.0×105 AU/mL): C/N=0.17, C/P=0.69, N/P=4.17 (C=sucrose, N=asparagine, P=phosphate, T=Tween-80).