[Objective] Research on the fermentation medium to produce low-temperature lipase strain CZW001. [Methods] Based on single factor test results, the liquid fermentation medium for the production of cold-active lipase by CZW001 was optimized using Plackett-Burman, Box-Behnken and response surface method under follow conditions: temperature 20 °C, pH 8.0, stirred velocity 160 r/min and fermentation time 2 d. [Results] The optimal fermentation medium of the strain is as follows (g/L): glucose 7.68, olive oil 21.93, (NH4)2SO4 2.0, KH2PO4 1.0, MgSO4·7H2O 0.27, CaCl2 0.3, NaCl 20.0, Tween-80 1.0. Under these conditions, the highest total enzyme activity (62.8 U/mL) was obtained and increased by 3.14 times compared with the original value before optimization. [Conclusion] The study on optimizing the fermentation medium to produce low-temperature lipase strain CZW001 obviously improved the lipase enzyme activity at low temperature.