Highly efficient production of monoglycerides by the continuous removal of fatty acids from lipase-catalyzed oil hydrolysis

Highly efficient production of monoglycerides was achieved from lipase-catalyzed oil hydrolysis by the continuous addition of CaCl₂ to remove the fatty acids produced. A fusion protein produced by connecting a cellulose-binding domain of Trichoderma hazianum cellulase to Bacillus stearothermophilus L1 lipase was used as a model 1,3-regiospecific lipase. The reaction was performed at pH 10 and 50°C, and the relationship between continuous removal of fatty acids and the production of monoglyceride was investigated by microscopic and HPLC analysis of oil emulsions and the reaction products. Without the addition of Ca²⁺ the reaction was inhibited by fatty acids, with the decrease in reaction rate being proportional to the concentration of fatty acids. When CaCl2 was continuously added in a 1:2 molar ratio with the released fatty acids, the reaction progressed unimpeded due to the formation of Ca-soaps. Both the yield and the fraction of monoglyceride in the reaction product increased due to the continuous removal of fatty acids.