Lyso-phospholipids enriched in n-3 polyunsaturated fatty acids have both surface/interface-active functionality and health-beneficial fragments such eicosapentaenoic acid (EPA, C20:5ω-3) and docosahexaenoic acid (DHA, C22:6ω-3); which could address multi-function demands in different fields. Starting with mariane phospholipids, we developed an entirely green process for enzymatic production of lyso-phosphatidylcholine. A few liquid lipases are screened for ethanolysis of purified phosphatidylcholine (PC), TLC-FID and GC-FID detections were used to monitor reaction progress and examine fatty acid compositions of different product fractions. Lipase NS-40116 from a genetic C. antarctica lipase A (CAL-A) displayed the best performance in either yield of lyso-PC and n-3 PUFA enrichment. Critical parameters, such as ethanol:PC ratio, reaction time and temperature, water content, and enzyme dosage are optimized. The results showed that, under optimized conditions through just one step-process, the yield of lyso-PC is up to 85% and the total n-3 PUFAs enriched up to 90% and DHA up to 75% in lyso-PC fraction. 31P/13C NMR analyses were used to illustrate the reaction mechanism, which verified that the regioselectivity and high non-hydrolytic property towards long chain PUFAs of CAL-A are the main character to drive this high efficiency and high selective process.