Selecting effective dietary strategies for professional football players requires comprehensive information on their energy expenditure (EE) and dietary intake. This observational study aimed to assess EE and dietary intake over a 14-day period in a representative group (n = 41) of professional football players playing in the Dutch Premier League (Eredivisie). Daily EE, as assessed by doubly labelled water, was 13.8 ± 1.5 MJ/day, representing a physical activity level (PAL) of 1.75 ± 0.13. Weighted mean energy intake (EI), as assessed by three face-to-face 24-h recalls, was 11.1 ± 2.9 MJ/day, indicating 18 ± 15% underreporting of EI. Daily EI was higher on match days (13.1 ± 4.1 MJ) compared with training (11.1 ± 3.4 MJ; P < 0.01) and rest days (10.5 ± 3.1 MJ; P < 0.001). Daily carbohydrate intake was significantly higher during match days (5.1 ± 1.7 g/kg body mass (BM)) compared with training (3.9 ± 1.5 g/kg BM; P < 0.001) and rest days (3.7 ± 1.4 g/kg BM; P < 0.001). Weighted mean protein intake was 1.7 ± 0.5 g/kg BM. Daytime distribution of protein intake was skewed, with lowest intakes at breakfast and highest at dinner. In conclusion, daily EE and PAL of professional football players are modest. Daily carbohydrate intake should be increased to maximize performance and recovery. Daily protein intake seems more than adequate, but could be distributed more evenly throughout the day.