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In recent years, probiotics of human origin have shown superior results and performance compared to probiotics from plant or dairy sources, in both in vitro and animal studies. Towards this end, the current study was conducted to explore the ergogenic properties of <i>Bifidobacterium longum subsp. longum</i> OLP-01 isolated from the intestinal microbiome of the gold medalist from the 2008 Beijing Olympics women&#8217;s 48 kg weightlifting competition. Male Institute of Cancer Research (ICR) mice were divided into four groups (<i>n</i> = 10 per group) and orally administered OLP-01 for 4 weeks at 0 (vehicle), 2.05 &#215; 10⁹ (OLP-01-1X), 4.10 &#215; 10⁹ (OLP-01-2X), and 1.03 &#215; 10¹⁰ (OLP-01-5X) CFU/kg/day. Physical performance tests including grip strength and endurance time were measured, with OLP-01 supplementation dose-dependently elevating grip strength and endurance. The anti-fatigue activity levels of serum lactate, ammonia, glucose, blood urea nitrogen (BUN), and creatine kinase (CK) were measured after an acute exercise challenge, and OLP-01 was found to significantly decrease lactate, ammonia, and CK levels. OLP-01 treatment was also found to significantly increase the resting levels of both hepatic and muscular glycogen, an indicator of energy storage. Supplementation by OLP-01 showed no subchronic toxic effects while supporting many health-promoting, performance-improving, and fatigue-ameliorating functions.