Characterizing the effect of limited oxygen availability on brain metabolism during brain activation is an essential step towards a better understanding of brain homeostasis and has obvious clinical implications. However, how the cerebral oxygen extraction fraction (OEF) depends on oxygen availability during brain activation remains unclear, which is mostly attributable to the scarcity and safety of measurement techniques. Recently, a magnetic resonance imaging (MRI) method that enables noninvasive and dynamic measurement of the OEF has been developed and confirmed to be applicable to functional MRI studies. Using this novel method, the present study investigated the motor-evoked OEF response in both normoxia (21% O2) and hypoxia (12% O2). Our results showed that OEF activation decreased in the brain areas involved in motor task execution. Decreases in the motor-evoked OEF response were greater under hypoxia (−21.7% ± 5.5%) than under normoxia (−11.8% ± 3.7%) and showed a substantial decrease as a function of arterial oxygen saturation. These findings suggest a different relationship between oxygen delivery and consumption during hypoxia compared to normoxia. This methodology may provide a new perspective on the effects of mild hypoxia on brain function. © The Author(s) 2021.