Abstract: Phytosterol and fatty acid were isolated from soybean oil deodorizer distillate respectively. The extracting conditions of phytosterol were optimized and the optimal conditions were as follows:the ratio of methanol to acetone was 1:2 (V/V);the ratio of solvent to sample was 3:1 (V/W);n-hexane was selected as detergent.The isolated fatty acid mainly included oleic acid and linoleic acid. CRL lipase was immobilized by physical adsorption and immobilization degree and enzyme activity were determined as selecting indexes. HP20 resin was determined as the most suitable carrier because of its good performance. The synthesis of phytosterol fatty acid ester was carried out by the obtained phytosterol and fatty acid under the catalysis of immobilized enzyme, which provides a new pathway for the practice of all kinds of areas.