To investigate sex differences in the organization of enzyme activities of energy supplying metabolism in skeletal muscle, samples of the vastus lateralis were extracted from active but untrained males (n = 16) and females (n = 17), ranging in age from 18 to 22 years. Muscle tissue from 2 different biopsy samples from each subject were analyzed for enzymes representative of the citric acid cycle (succinic dehydrogenase, SDH), beta-oxidation of fatty acids (3-hydroxyacyl CoA dehydrogenase, HAD), glycogenolysis (phosphorylase, PHOSPH), glycolysis (pyruvate kinase, PK; phosphofructokinase, PFK and lactate dehydrogenase, LDH) and glucose phosphorylation (hexokinase, HK). The results indicated that the maximal activities of PFK, PK, LDH and PHOSPH, HK and SDH averaged between 15 and 32% higher in the males than in the females. No significant differences between the sexes were found for HAD. When enzyme activity ratios were calculated, sex differences were only evident for the HAD/SDH ratio (mean +/- SD; females = 0.56 +/- 0.20; males = 0.41 +/- 0.11 and for the PFK/HAD ratio (females = 7.40 +/- 1.6; males = 9.58 +/- 1.9). The findings suggest that (1) the females have a significantly lower overall capacity for aerobic oxidation and for anaerobic glycolysis than the males; (2) the females have a greater capacity for beta-oxidation relative to the capacity of the citric acid cycle; and (3) the glycolytic potential relative to the potential for beta-oxidation is lower in the females.