The optimal conditions of the downstream process for recovery of pullulan produced by Aureobasidium pullulans HP-2001 were examined using response surface method (RSM). The optimal amount of diatomite in filter press and the optimal flow rate in a continuous flow centrifuge for removal of cells from the culture broth of A. pullulans HP-2001 were found to be 5.0% (v/v) and 2.0 L/min. Based on central composite design (CCD) experiments and analysis of variance (ANOVA) indicated that the optimal conditions for recovery of pullulan from the supernatant by precipitation were the volume ratio of ethanol (or isopropanol) to supernatant of 3.0 : 1.0, the reaction time of 29.5 h, and the reaction temperature of 20.2 ℃. The expected maximal recovery yields of pullulan using ethanol and isopropanol under optimized conditions were 79.2 and 85.5%, respectively.