A rotational reciprocating plate impeller prototype, designed to improve the mixing homogeneity of viscous non-Newtonian fermentation broth, has been tested in pullulan fermentations. With this new impeller, the operating levels of several factors were investigated to improve pullulan production with Aureobasidium pullulans ATCC 42023 in a 22-L bioreactor using experimental designs. Because both high molecular weight (MW) and high concentration of pullulan were desired; the exopolysaccharide (EPS) concentration and the broth viscosity were used as optimization objective functions to be maximized. A 6-run uniform design was used to investigate five factors. Under the best operating conditions among the six runs, 29.0 g L(-1) EPS was produced at 102 h. This condition was used as the starting point for further investigation on the two statistically significant factors, the pH and the agitation speed. An 8-run 3-level custom design that investigates up to second-order effects was used in the second stage. An optimal zone of operating conditions for large quantity of high MW pullulan production was identified. A concentration of 23.3 g L(-1) EPS was produced at 78 h. This is equivalent to an EPS productivity of 0.30 g L(-1) h(-1). The corresponding apparent viscosity of the broth was 0.38 Pa s at the shear rate of 10 s(-1).