Immobilization of phospholipase C for the production of ceramide from sphingomyelin hydrolysis

The immobilization of Clostridium perfringens phospholipase C was studied for the first time and the catalytic properties of the immobilized enzyme were investigated for the hydrolysis of sphingomyelin to produce ceramide. Ceramide is of great commercial potentials in cosmetic and pharmaceutical industries such as in hair and skin care products, due to its major role in maintaining the water-retaining properties of the epidermis. The feasibility of enzymatic production of ceramide through hydrolysis of sphingomyelin has previously been proven. In order to improve the reusability of the enzyme, the present study was focused on the immobilization of phospholipase C in the production of ceramide from sphingomyelin. By screening nine different carriers, the enzyme immobilized on Lewatit had the highest catalytic activity towards sphingomyelin hydrolysis. Pre-wetting Lewatit with ethanol leaded to higher enzyme fixation on the carrier, but the activity of the enzyme was decreased. Increasing the initial enzyme concentration resulted in more enzyme adsorption on the carrier, where the specific activity was increased. Through optimization of the reaction using the immobilized enzyme, the optimal temperature was around 46 oC and optimal water volume was 3.5 %. The reaction had little dependence on pH. After 7 recycles, immobilized enzyme retained around 70 % of the initial activity. Immobilized enzyme was deactivated irregularly when stored at room temperature, but followed first order deactivation when stored at 40 oC.