Modeling and optimization of lipase-catalyzed synthesis of phytosteryl esters of oleic acid by response surface methodology

Enzymatic esterification of phytosterols with oleic acid to produce phytosteryl esters was performed in hexane. Response surface methodology was used to model the reaction. Candida rugosa lipase was the biocatalyst for the reaction. The reaction factors investigated were temperature (Te = 35–55 °C), reaction time (t = 4–24 h), substrate molar ratio (Sr = 1–3, oleic acid:phytosterols), and enzyme amount (En = 2–10%). Well-fitting quadratic polynomial regression model for degree of esterification (DE) was established after regression analysis with backward elimination and verified by a χ2 test. All factors investigated positively affected DE, with t having the greatest effect followed by En, Sr, and Te. The quadratic terms of t, Sr, and En showed negative effects on DE, whereas, that of Te had no effect on DE. Optimal reaction conditions were: Te, 51.3 °C; t, 17.0 h; Sr, 2.1; En, 7.2% and DE was 97.0 mol% under these conditions.