The objective of the present study was to determine the dose response of the methane (CH4) inhibitor 3-nitrooxypropanol (NOP) on enteric CH4 production and dry matter intake (DMI) for beef cattle fed a high-forage or high-grain diet. Fifteen crossbred yearling steers were used in two consecutive studies (high-forage backgrounding, high-grain finishing), each designed as an incomplete block with two 28-day periods with a 7-day washout in between and treatments corresponding to six doses of NOP (0 (Control), 50, 75, 100, 150, 200 mg/kg DM). The NOP was provided in the ration daily with the dose increased gradually over the first 10 days of each period. No treatment effects were observed on overall DMI or DMI of cattle when they were in the chambers either for the high-forage (P ≥ 0.54) or high-grain (P ≥ 0.26) diet. With the high-forage diet, NOP supplementation lowered total CH4 emissions (g/day) (P = 0.05), with the response at 200 mg NOP/kg DM different from Control (P < 0.05). Similarly, CH4 emissions corrected for DMI (g/kg DMI) and as a percentage of gross energy intake were linearly reduced in the high-forage diet with supplemental NOP (P < 0.01) and responses observed at 100, 150 and 200 mg NOP/kg DM differed from Control (P < 0.05). For the high-grain diet, total CH4 emissions decreased with incremental increases in the concentration of NOP supplemented (P = 0.04) and responses observed at 150 and 200 mg/kg DM differed from Control. Similarly, linear responses were observed with CH4 emissions corrected for DMI (P = 0.04) and gross energy intake (P = 0.02), with 100–200 mg NOP/kg DM differing from Control. Overall, results from the present study demonstrated that for beef cattle fed high-forage and high-grain diets, supplementation of 100–200 mg NOP/kg DM lowered enteric CH4 emissions without inducing any negative effects on DMI.