Characterization of the recombinant porcine pancreas phospholipase A2 expressed in Pichia pastoris GS115 and its application to synthesis of 2-DHA-PS

Abstract

1-Acyl-2-docosahexaenoyl-phosphatidylserine (2-DHA-PS) is a species of phosphatidylserine (PS) with a docosahexaenoic acid (DHA) esterified sn-2 position. 2-DHA-PS has been suggested to enhance cognitive performance. Enzyme-catalyzed synthesis of 2-DHA-PS is proceeded by using phospholipase A2. The codon-optimized gene pla2m, encoding porcine pancreas phospholipase A2 (ppPLA2), was expressed in Pichia pastoris GS115 for functional characterization of recombinant PLA2M (rPLA2M). The rPLA2M showed maximum enzymatic activity at 40°C and pH 8.0 and was stable within a broad range of temperatures (30–55°C) and pHs (pH 6.0-9.0). Moreover, rPLA2M was successfully applied in the synthesis of 2-DHA-PS using PS and DHA as substrates with a yield of 23%. Thus, rPLA2M displays the same hydrolysis and transesterification activities as native ppPLA2, providing a novel strategy for the production of 2-DHA-PS.