BACKGROUND: Methane emission from enteric rumen fermentation is a main source of greenhouse gas (GHG) emission and a major concern for global warming. RESULTS: In this study, we isolated methanotroph-methylotroph consortium NC52PC from the rumen after a series of sub-culture and repetitive streaking on an agar plate and polycarbonate membrane filter. The NC52PC comprises methanotroph species (Methylocystis sp.) and methylotroph species (Methylobacterium sp.), forming a consortium capable of growing solely on methane as a carbon source. Their morphology, growth, and genome sequence were characterized. We assessed its effectiveness in mitigating methane emissions through both in vitro and in vivo experiments. During the in vitro trial, the introduction of NC52PC (at a concentration of 5.1 × 10 CONCLUSION: Feeding methanotroph-methylotroph consortium NC52PC significantly reduced methane emissions in Korean beef cattle without any adverse effects on animal health. These findings suggest that this probiotic could serve as a promising feed additive to effectively mitigate methane emissions from ruminants. However, further research is needed to evaluate the long-term effects of NC52PC on animal health, and on meat and milk quality.