Production and Scale-up of phosphatidyl-tyrosol catalyzed by a food grade phospholipase D

Highly purified phosphatidyl-tyrosol was obtained by using a food grade phospholipase D from Actinamadure sp. transphosphatidylation in a GRAS (Generally Recognized As Safe) biphasic medium. The reaction medium, comprised of an aqueous phase and ethyl butyrate, has been considered as an alternative to other biphasic systems previously reported utilizing more harmful organic solvents. The purpose of the present study was to purify phosphatidyl-tyrosol from a transphosphatidylation reaction mixture by using a procedure readily scalable to obtain a new valuable food ingredient. Initially, phosphatidyl-tyrosol was purified via semi-preparative HPLC equipment to be used as analytical standard. The best results of the transphosphatidylation reaction were obtained for two different PC concentrations, namely 83 and 166 mmol/L, with PC conversion of ca. 97 and 94% (w/w) and a final phosphatidyl-tyrosol concentration of 81 and 157 mmol/L, respectively. Finally, the procedure was scaled-up and 40 g of highly purified phosphatidyl-tyrosol (97% (w/w)) were readily purified by centrifugation without involving the utilization of organic solvents.

► Phosphatidyl-tyrosol by a food grade phospholipase D has been produced.
► GRAS biphasic medium has been proposed as an alternative to other biphasic systems.
► Substrate ratios, concentrations, enzyme loading and reuse have been revised.
► A readily scalable purification method for phosphatidyl-tyrosol has been developed.
► 40 g of purified phosphatidyl-tyrosol were finally isolated by a simple procedure.