With an aim to explore the effects of β‐alanine (β‐A) on spatial memory and fatigue resistance, Kunming mice were treated with different concentrations of β‐A (418, 836, and 2090 mg·kg^‐1 ·day^‐1). After gavage feeding with β‐A for 10 weeks, results of the maze and MWM tests showed that β‐A can enhance spatial learning and memory in mice. After evaluating the fatigue resistance, biochemical parameters (LG, GG, BUN, SOD, and MDA) showed significant differences in the low concentration treatment group compared to control group. Our data demonstrated that the appropriate dose of β‐A can alleviate the oxidative stress and muscle fatigue in mice. Subsequently, expression of mRNA of key genes involved in cAMP‐PKA pathway (PDE4A, MAPK1, adcy1, cAMP and CREB) was up regulated. Also, expression levels of apoptotic pathway genes were significantly affected as confirmed by qPCR and Western blotting. Our results demonstrated that β‐A can enhance spatial learning and memory in mice via regulation of cAMP‐PKA and apoptotic pathway.