Taking the conversion rate of sterols as a indicator, the effects of 10 common commercial lipases in catalyzing the synthesis of lignosterol oleate were investigated, and the Candida rugosa lipase was determined as the preferred biocatalyst, and n-hexane was further screened out as the preferred reaction medium. Based on the investigation of the four reaction conditions of lipase dosage, oleic acid-alcohol molar ratio, reaction temperature and reaction time, the process conditions of enzyme-catalyzed lignosterol oleate synthesis were optimized by response surface analysis. The optimized process parameters of Candida rugosa lipase catalyzed synthesis of lignosterol oleate are: CRL addition amount is 10 w% based on the sterols, the molar ratio of substrate acid to alcohol is 3.8:1, the reaction temperature is 46 ℃, the reaction time is 28 h, and the conversion rate of sterol is 91.56%.