A comprehensive approach, based on the use of , mouse, and human models, elucidates the impact of TWK10 on exercise performance and longevity.

The functionality of probiotics is highly influenced by culture and processing conditions, making batch stability validation through human or mouse trials impractical. Here, we employed a comprehensive approach using , mouse and human models to elucidate the beneficial effects of TWK10 (TWK10). In , TWK10 administration significantly prolonged lifespan by 26.1 ± 11.9 % ( < 0.05), enhanced locomotion ( < 0.01) and muscle mass ( < 0.001), elevated glycogen storage ( < 0.05), and reduced lipid accumulation ( < 0.001), outperforming GG and type strain ATCC 14917. We also confirmed the equivalence of laboratory-prepared and mass-produced TWK10 in ergogenic efficacy using assay. In mice, oral administration of mass-produced TWK10 significantly enhanced exercise performance and glycogen storage in muscle and liver in a dose-dependent manner. In a clinical study involving healthy male adults, significant improvements in grip strength (1.1-fold,  < 0.01) and exhaustion time (1.27-fold,  < 0.01), and significant reductions in circulating lactate and ammonia levels were observed in the TWK10 group (1 × 10 colony-forming unit/day) compared to the control group. Both humans and mice receiving mass-produced TWK10 showed improved body composition with increased muscle mass and reduced fat mass. In conclusion, TWK10 demonstrates superior longevous and ergogenic effects in compared to reference strains. The consistent ergogenic efficacy of mass-produced TWK10 across , mice, and humans, highlights the utility of as a reliable model for probiotic research and industrial application.