The physicochemical properties of wheat bran have an effect on its technofunctional and nutritional profile. The possibility to induce physicochemical modifications in wheat bran using microfluidisation was investigated. An I-optimal experimental design was used to investigate the effect of microfluidisation processing parameters (pressure, number of passes, bran concentration and initial particle size) on important properties of wheat bran (particle size, microstructure, chemical composition, water retention capacity (WRC), extractability, viscosity and sedimentation). With the parameters used in this study, microfluidisation reduced wheat bran median particle size to 14.8 μm and disintegrated starch granules from the attached endosperm. This coincided with an increased extractability of starch and arabinoxylan. While the initial particle size was of minor importance, a higher pressure, larger number of passes and lower bran concentration during microfluidisation resulted in a smaller particle size, higher WRC and extractability, and an increased viscosity and stability in a 2% wheat bran suspension.