The invention provides an enzymatic preparation method of phosphatide-type DHA. The enzymatic preparation method comprises the following steps: DHA production bacteria are cultured, bacterium bodies are obtained, the following enzyme reaction system is added into a three-necked flask, specifically, granulesten and 2.5 mL-5 mL of DHA grease are added simultaneously, and immobilized lipase is addedinto a mixture; thermostatic water bathing is conducted, the immobilized lipase is filtered and recovered, and filtrate is obtained; the filtrate is subjected to rotary evaporation at 45 DEG C-60 DEGC to be in a thick oil shape, and an oil phase is obtained; acetone is added into the oil phase, centrifuging is conducted, and sediments are collected; and after the acetone is blow-dried through nitrogen, freeze drying is conducted, and the phosphatide-type DHA is obtained. The invention provides an in-vitro enzymatic catalysis conversion technology, and in-vitro conversion preparation of the phosphatide-type DHA can be quickly, safely and efficiently achieved.