Synthesis of diacylglycerols containing CLA by lipase-catalyzed esterification

Diacylglycerols (DAG) were prepared by esterification of glycerol with conjugated linoleic acid (CLA) in the presence of an immobilized 1,3-regiospecific lipase from Rhizomucor miehei and vacuum conditions. The effects of several parameters, namely, temperature, enzyme loading, stirring speed, and vacuum, on the concentration and the purity of the DAG were studied. The reaction temperature influenced both the reaction rate and the concentration of the DAG. The rate of DAG synthesis increased as the enzyme loading increased. However, for high enzyme loadings, the concentration of triacylglycerols (TAG) increased significantly at long reaction times and, as a result, the purity of the DAG decreased. When the stirring speed increased from 150 to 450 rpm, the DAG concentration increased significantly. However, at stirring speeds above 450 rpm, no significant increases in DAG concentration were observed. When the pressure was decreased from 20 to 3 mmHg, the maximum concentration of DAG increased from 76.0% to 80.5%. No increase in the DAG concentration was observed when the pressure was decreased from 3 to 1 mm Hg, even though a slightly higher DAG purity was achieved at 1 mm Hg. For the range of absolute pressures tested, the concentrations of 1,2-DAG were less than 1%.