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Response surface methodology (RSM) was taken to enhance the production of low-temperature-active phytase from the strain Kurthia CZC0806 which was isolated from the Yellow Sea in China. The Plackett-Burman and Box-Behnken design were applied to optimize the fermentation medium components of the strain. Peptone, saccharose and NaCl were selected as the most significant factors, based on the Plackett-Burman design. The Box-Behnken was designed for these three factors. The results were analyzed by using the Minitabl5.0 soft. The optimal medium is: Peptone, saccharose and NaCl (at 20°C, initial pH 7.0). In the optimal medium conditions, the activity of the enzyme is 103.11 U/mL.