The effects of DO and pH on the mass production of pullulan with high molecular weight and the morphology of A. pullulans ATCC 42023 were evaluated. A. pullulans showed a maximum production of pullulan (11.98 g/l) when the initial pH of the culture broth was 6.5 in a shake-flask culture. In a batch culture, the mixture of a yeast-like and mycelial cell forms was found at a pH of 4.5, and the maximum production of pullulan (13.31 g/l) was obtained. However, a high proportion of high molecular weight pullulan (M.W. >2,000,000) was produced at a pH of 6.5, with a yeast-like morphology. The maximum pullulan production yield (51%) was obtained at a pH noncontrol (initial pH 6.5) and DO control (above 50%) condition. Pullulan degrading enzyme was activated when the pH of the broth was lower than 5.0 and the portion of low molecular weight pullulan was increased. The formation of a black pigment was observed at an initial stationary phase, at 40 h of fermentation. Therefore, the fermentation should be carried out in a pH noncontrol (initial pH of 6.5) and DO control (above 50%) condition, and should be harvested before reaching the stationary phase (around 40 h) for the production of high molecular weight pullulan.