Sterol ester production using lipase-catalyzed reactions in supercritical carbon dioxide

Synthesis of sterol and stanol esters is of importance, due to their recent recognition and application in the food and nutriceutical industries as cholesterol-lowering agents. In this study, several enzymes were evaluated to determine the best catalyst and optimal conditions for the reaction between various fatty acids and cholesterol or sitostanol in supercritical carbon dioxide (SC-CO2). Using an analytical supercritical fluid extraction (SFE) unit, the lipase derived from Burkholderia cepacia, Chirazyme L-1, was determined to be the most selective for facilitating the desired reactions. Fatty acids C-8-C-18, pressures between 20.7 MPa and 31 MPa, a temperature range of 40-60 degreesC, along with variable flow rates, and initial static hold times were used to evaluated the feasibility of the above reaction. The yield of the cholesterol esters, as measured by supercritical fluid chromatography (SFC), ranged from 90% for caprylic acid to 99% for palmitic acid, while the corresponding reaction between sitostanol and the same fatty acids produced yields of 92% for caprylic acid and 99% for palmitic acid, respectively. The extraction apparatus was modified to provide a continuous flow of the reagent fatty acid and sterol/stanol over the enzyme bed, thereby allowing continuous production of the desired esters which averaged a 99% yield under optimal conditions.