摘要:The current work aimed to evaluate the effect of ultrasonic treatment on the enzymatic transesterification of medium–long–medium (MLM) lipids using 2‐monoacylglycerol, bearing distinct fatty acids at the sn ‐2 position with palmitic acid, octadecanoic acid, oleic acid, eicosapentaenoic acid, and docosahexaenoic acids as sn ‐2 acyl donors. The effects of ultrasonic treatment conditions, including substrate concentration, reaction temperature and time, and enzyme loading, on the insertion of fatty acids into the sn ‐2 acyl position of MLM lipids were investigated. The data showed that low‐frequency ultrasonic treatment could remarkably improve the insertion rate of polyunsaturated fatty acid (PUFA) into the sn ‐2 position of MLM lipids, compared with the conventional treatment method. By increasing the ultrasonic frequency from 20 to 30 KHz, while maintaining power at 150 W, the rate of synthesis of monounsaturated fatty acid and PUFA increased from 23.7% and 26.8% to 26.6% and 32.4% ( p < 0.05), respectively. Moreover, ultrasonic treatment reduced the optimum reaction temperature from 45 to 35°C. However, the activity of Lipozyme RM‐IM treated with ultrasound considerably declined from 31.10% to 26.90% ( p < 0.05) after its fourth cycle, which was lower than that without ultrasonic treatment. This work provokes new routes for the utilization of ultrasonic technology in the synthesis of MLM lipids using different fatty acids as sn ‐2 acyl donors.