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

1-Acy1-2-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 A(2). The codon-optimized gene pla(2)m, encoding porcine pancreas phospholipase A(2) (ppPLA(2)), was expressed in Pichia pastoris GS115 for functional characterization of recombinant PLA(2)M (rPLA(2)M). The rPLA(2)M showed maximum enzymatic activity at 40 degrees C and pH 8.0 and was stable within a broad range of temperatures (30-55 degrees C) and pHs (pH 6.0-9.0). Moreover, rPLA(2)M was successfully applied in the synthesis of 2-DHA-PS using PS and DHA as substrates with a yield of 23%. Thus, rPLA(2)M displays the same hydrolysis and transesterification activities as native ppPLA(2), providing a novel strategy for the production of 2-DHA-PS. (C) 2016 Elsevier Ltd. All rights reserved.