The objectives of the two experiments presented were to determine the separate effects of diet and mobility associated with grazing on the metabolic potential of muscles of Charolais steers reared at similar growth rates and slaughtered at the same age (31 months on average). Glycolytic [lactate dehydrogenase (LDH), phosphofructokinase (PFK)] and oxidative [β-hydroxyacyl-CoA dehydrogenase (HAD), citrate synthase (CS), isocitrate dehydrogenase (ICDH), cytochrome c oxidase (COX)] enzyme activities were measured spectrophotometrically in semitendinosus (ST), a glycolytic muscle, and rectus abdominis (RA), an oxidative muscle. In Experiment 1, 24 weaned steers were assigned to two groups: grazing grass (GG) and maize silage (MS). In Experiment 2, 30 weaned steers were assigned to five groups: grazing grass (GG), cut grass without walking (CG), cut grass with walking (CGW), maize silage without walking (MS) and maize silage with walking (MSW). Both experiments lasted for 2 years. All grass fed animals were fed grass silage during the two winter seasons. During the two summer seasons, GG steers were grazing and did not receive any supplementation while CG and CGW steers were fed indoor freshly cut grass alone. MS and MSW steers were fed maize silage indoor during the entire experiment. For the second summer before slaughter, half of the indoor fed steers on each type of diet (CGW and MSW groups) was submitted 7 days a week to a 5.2 km walk daily. At slaughter, carcass weight and composition did not differ between groups. Oxidative HAD, CS and ICDH enzyme activities were higher for GG, as compared with MS group, in RA muscle in both experiments (P < 0.05 to 0.01) and in ST muscle (P < 0.05) in Experiment 2. Muscles from the GG group were thus more oxidative than those of the MS group. It was further tested whether these effects could be specifically attributed to the nature of the diet or to differences in mobility. A cut grass diet (vs. maize silage) increased HAD and ICDH activities (P < 0.05) and mobility induced by walking increased HAD and CS (P < 0.01) activities. In conclusion, the more oxidative metabolic orientation of muscles of grazing steers originates from a combination of two effects: an increase mobility at pasture and a grass (vs. maize silage)-based diet.