The aim of the study was to evaluate the effect of crude protein (CP) levels in concentrate and Sesbania grandiflora pod meal (SG) supplementations on feed intake, rumen fermentation, and methane (CH4) mitigation in Thai purebred beef cattle. Four cattle with 100 ± 5.0 kg body weight were used in this study. A 2 × 2 factorial experiment in a 4 × 4 Latin square design were conducted, in which factor A was the CP contents in concentrate of 14, and 16% of dry matter (DM) and factor B was the supplement contents of SG at 0.4% and 0.6% DM intake, respectively. The results showed that the CP contents in concentrate and SG had no interaction effect on intake, digestibility, ruminal ecologies, ruminal fermentation products, and nitrogen utilization. Increasing CP contents in concentrate did not influence DM intake and nutrients’ digestibility, and SG supplementation at 0.6% significantly (P<0.05) decreased CP digestibility. Increasing CP content to 16% increased significantly (P<0.05) the ruminal ammonia nitrogen (NH3-N) concentration while decreased significantly (P<0.05) the NH3-N concentration, protozoal number, and blood urea nitrogen (BUN) at 4 h post-feeding. The 0.6% supplementation of the SG increased significantly average total volatile fatty acids (VFAs) and propionate (C3) concentration while decreased significantly average acetate (C2), C2:C3 ratio, and CH4 production, which was 2.71% for C2, 13.17% for C2:C3 ratio, and 4.37% for CH4 production lower than 0.4% supplementation. Fecal nitrogen excretion was significantly decreased when supplemented with 0.6% of the SG. In conclusion, 0.6% of the SG supplementation showed a greater effect on intake, rumen manipulation, and CH4 mitigation and would recommend supplementation to a concentrate-based diet containing either 14% or 16% CP content.