Technology conditions for diacylglycerol preparation from rice bran oil were optimized by response surface methodology. On the basis of four single factor experiments, for reaction temperature, reaction time, immobilized lipase addition quantity and reactants weight ratio were chosen as influence factors, through four-factor quadric orthogonal regression test and structure matrix constructing, quadratic regression model with diacylglycerol yield as response value was set up and analyzed. The results showed that optimal preparation conditions were reaction temperature 26 ℃, reaction time 10 h, immobilized lipase addition 8.5% and reactant weight ratio 14∶1. Under these optimized conditions, the theoretical yield was 60.36%, practical yield was 58.31% with relative error 3.4%.