The design of an optimum and cost-efficient medium for production of cold-active cellulase by Penicillium cordubense D28 was attempted by using response surface methodology (RSM). Based on the Plackett–Burman design, corn meal, (NH4)2SO4 and branc were selected as the most critical nutrient. Subsequently, they were investigated by the Box-Behnken design. Results showed that the maximum cold-active cellulase activity of 110.4U/mL was predicted when the concentration of corn meal, (NH4)2SO4 and branc were 21.97 g/L, 2.39 g/L and 14.99 g/L, respectively. The results were further verified by triplicate experiments. The batch reactors were operated under an optimized condition of the respective corn meal, (NH4)2SO4 and branc concentration of 22 g/L , 2.4 g/L and 15 g/L , the initial pH of 6.0 and experimental temperature of 20 ± 1°C. Without further pH adjustment, the maximum cold-active cellulase activity of 109.8 U/mL was obtained based on the optimized medium with further verified the practicability of this optimum strategy.