Partitioning of evapotranspiration (ET) into transpiration (T) and residual evaporation (E) is a challenging but important task in order to assess the dynamics of increasingly scarce water resources in forest ecosystems. The T/ET ratio has been linked to the ecosystem water use efficiency of temperate forests, and thus is an important index for understanding utilization of water resources under global climate change. We used concurrent sap flow and eddy-covariance measurements to quantify the ET partitioning in pure European beech forest during the 2019–2020 period. The sap flow data were upscaled to stand level T and combined with stand level ET to calculate the T/ET ratio. We analysed intra-annual dynamics, the effect of seasonality and the impact of meteorological conditions on T, ET and T/ET. Annual T/ET of a pure European beech ecosystem was 0.48, falling at the lower end of reported global T/ET values for forest ecosystems. T/ET showed significant seasonal differences throughout spring (T/ET = 0.28), summer (T/ET = 0.62) and autumn (T/ET = 0.35). Air temperature (R2 = 0.45–0.63), VPD (R2 = 0.47–0.6) and PAR (R2 = 0.32–0.63) affected the daily dynamics of T, ET and T/ET; however, soil water content (SWC) had no significant effect. Mature European beech trees showed more anisohydric behaviour and relatively stable T/ET, even under decreasing SWC. The results improve the understanding of ecosystem scale T, ET and T/ET intra-annual dynamics and environmental constraints in anisohydric mature European beech.