Knowledge and prediction of mixing and mass transfer in agitated bioreactors is fundamental for process development and scale up. In particular key process parameters such as mixing time and volumetric mass transfer coefficient are essential for bioprocess development. In this work the mixing and mass transfer performance of a high power agitated pilot scale bioreactdr has been characterized using a novel combination of computational fluid dynamics (CFD) and experimental investigations. The effect of turbulence inside the vessel was predicted using a standard RANS k-s model. Mixing time was investigated by carrying out sodium chloride tracer experiments for both Newtonian and non-Newtonian fluids at various viscosities and agitation speeds, while tracking the conductivity. The mixing performance was simulated with CFD and the results showed good agreement with the experimental data. The mass transfer coefficients were determined from six Trichoderma reesei fermentations at different well-defined process conditions. Similarly the mass transfer was predicted by Higbie's penetration model from two-phase CFD simulations using a correlation of bubble size and power input, and the overall mass transfer coefficients were in accordance with the experimental data. This work illustrates the possibility of predicting the two phase fluid dynamic performance of an agitated pilot scale bioreactor using validated CFD models. These models can be applied to illustrate the effect of changing the physical process conditions. (C) 2017 Elsevier Ltd. All rights reserved.