AbstractA two-stage, pH- and temperature-controlled wheat bran fermentation method using Aureobasidium pullulans was investigated for feruloyl oligosaccharides (FOs) production and the activities of xylanase, xylosidase, and ferulic acid esterase (FAE). A. pullulans secreted xylanase, xylosidase, and FAE at high levels in the initial pH of 4.0 to 5.0 and a fermentation liquid temperature of 31 °C to 33 °C. FOs production via two-stage fermentation (FOs 2) reached 1123 nmol/L after fermentation for 96 h, by controlling the initial pH at 4.0 and the initial temperature at 33 °C, and then changing the pH to 6.0 and the temperature to 29 °C at the same time at 36 h. This process was 12 h shorter and 219 nmol/L higher than a one-stage fermentation for producing FOs 1. Xylanase, xylosidase, and FAE activities were highly correlated with controlled pH and temperature and FOs biosynthesis rate. Thus, the combination of two-stage controlled pH and temperature could support mass production of FOs.