期刊
PROCESS BIOCHEMISTRY
卷 51, 期 10, 页码 1472-1478出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.procbio.2016.06.023
关键词
Porcine pancreas phospholipase A(2); Pichia pastoris; Recombinant protein expression; 2-DHA-PS; Synthesis
资金
- National Natural Science Fund of China [31571805]
- Tianjin correspondent Program of Science and Technology of China [15JCIPJC56500]
- National High Tech Research and Development Plan of China [2013AA102106-07]
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.
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