A partial nitritation/anammox (PN/A) process was operated at two different mixing intensities to quantify the extent to which diffusional limitations impact process rates. At a steady-state operation, the total inorganic nitrogen removal efficiency in the bench-scale sequencing batch reactors was found to increase as mixing intensity decreased (62 and 84% for average velocity gradient (G) values of 15 and 5.3 s−1, respectively). The half-velocity constants (KO2) with respect to bulk-phase dissolved oxygen (DO) concentration for ammonia-oxidizing bacteria (AOB) and anaerobic ammonium-oxidizing (anammox) organisms were estimated on the basis of nitrogen removal rates that were observed in activity tests. The activity tests were conducted over a range of bulk-phase DO concentrations. The best-fit KO2AOB values were estimated to be 0.68 ± 0.34 and 0.54 ± 0.56 mg O2/L for G values of 15 and 5.3 s−1, respectively. The AOB values were not statistically different (p = 0.19) between mixing conditions which were consistent with AOB dominating the surface of granules. The best-fit KO2anammox values were estimated to be 0.13 ± 0.09 and 0.55 ± 0.40 mg O2/L for G values of 15 and 5.3 s−1, respectively, and were statistically different p = 3:9 × 10−10. The results demonstrated that mixing conditions should be considered when designing PN/A processes and provide quantitative results that can be employed to improve models of these processes.