Soil enzymes play critical roles in material cycle and energy flow of ecosystems. Understanding soil enzyme activities is of great significance for exploring ecosystem functions. In this study, we investigated soil enzyme activities, stoichiometry and their driving factors at six different altitudes (4300-5100 m) on Qinghai-Tibet Plateau alpine meadow using Biolog microplate analysis. The results showed that β-1,4-glucosidase (βG) closely related to C cycle, β-1,4-N-acetylglucosaminidase (NAG) and L-leucine aminopeptidase (LAP) closely related to N cycle and the activity of acid phosphatase (AP), which was closely related to P cycle, all exhibited unimodal trends with increasing altitude, with the order of 4800 m＞4950 m＞4400 m＞4650 m＞5100 m＞4300 m. Soil N:P enzyme activity ratio showed the same trend as soil enzyme activity, and reached the highest value at 4950 m, however, soil C:N and C:P enzyme activities ratios increased along the altitude. Pearson correlation analysis showed that SOC, TN and soil water content were significantly positively correlated with the activities of four types of enzymes. Mean annual precipitation was significantly negatively associated with the activities of NAG and AP. Mean annual precipitation, mean annual temperature, Shannon diversity, vegetation richness, vegetation coverage and TN affected ratios of soil C:P and N:P enzymes. Soil C:N activity ratio correlated with mean annual temperature, mean annual precipitation, vegetation richness, vegetation coverage, SOC and TN. In summary, soil enzyme activities and stoichiometry had remarkable difference along the altitude gradient on Qinghai-Tibet Plateau alpine meadow, with certain N limitation in high altitude areas. Soil water content, TN, SOC, mean annual precipitation and mean annual temperature were key factors driving such differences.