【Objective】Lipase-catalyzed synthesis of DAG production between perilla seed oil and glycerol was studied.【Method】Response surface methodology was used to optimize the reaction conditions of glycerolysis by Lipozyme RM IM in a solvent-free system. 【Result】 The optimal parameters were as follows: substrate molar ratio of glycerol to perilla seed oil was 1﹕1.5, enzyme load was 11.8 wt%, reaction temperature was 72℃, and reaction time was 14.5 h. Under the optimal conditions, DAG content could reach 54.33% in the reaction product, and TAG content lowered to 26.20%. The theoretical purity of DAG oil could reach 67.47% after purification by distillation, and the content of α-linolenic acid reached 52.26% in the DAG oil.【Conclusion】Substrate molar ratio of glycerol to perilla seed oil, enzyme load, reaction temperature and reaction time were all significant factors, and the interaction between substrate molar ratio and enzyme load was the most significant.