Abstract: Basa catfish oil with a high content of sn-2 palmitic acid and low content of total palmitic acid was a good raw material for preparation of 1,3-dioleoyl-2-palmitoylglycerol(OPO). In this study,Basa catfish oil was fractionated under 30 ℃ to enrich the fat fraction with higher content of sn-2 palmitic acid,and then the OPO product was prepared by acidolysis of the fat fraction with free fatty acids from high oleic acid sunflower oil using sn-1,3 specific lipase,Lipozyme RM IM,in a packed-bed reactor. The conditions of acidolysis reaction were optimized as follows residence time 1 h, substrate molar ratio 1 ∶ 6( fish oil / free fatty acid), reaction temperature 50 ℃,water content 3.5 wt%. Under these conditions,the product had 57.8% palmitic acid at sn-2position and 78.7% oleic acid at sn-1,3 positions.