Abstract: Phosphatidylserine (PS) is a new resource food, which has less content in nature and the difficulty of extraction. Phospholipase D (PLD) is the critical synthetase in enzymatic preparation of PS. This paper mainly introduces the characteristics of molecular structure of PLD and the catalytic mechanism and enzyme activity, and summarizes the current preparation methods of PS, focusing on the research progress and reaction system of enzymatic synthesis, and analyzes and prospects the selection of the reaction system for PS preparation. It is expected to provide reference for deepening the research of enzymatic preparation of PS, so as to promote the industrial production and application of new resource food PS.