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

- A novel strategy for synthesis of 1-acyl-2-docosahexaenoyl-phosphatidylserine (2-DHA-PS) using recombinant porcine pancreas phospholipase A2 (rPLA2M) is studied.
- rPLA2M is successfully expressed in Pichia pastoris GS115.
- rPLA2M displays the same activity as the native porcine pancreas phospholipase A2.
- rPLA2M is successfully applied in the synthesis of 2-DHA-PS.
- The yield of 2-DHA-PS using the rPLA2M is 23% in a safe method.

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.

139