Abstract: Urate oxidase is an enzyme that catalyzes the oxidation of uric acid to allantoin, which is widely used in pharmaceutical industry, treatment of gout, detection of blood uric acid concentration. It plays an important role in prevention and treatment of high uric acid hematic disease caused by tumor chemotherapy. Especially in recent years, high uric acid disease occurs so frequently that urate oxidase has attracted much more attention. A high-yielding urate oxidase strain was isolated and screened from the soil in Cangshan, Dali City, Yunnan Province. It was identified as a strain of Klebsiella by 16S rRNA gene sequence analysis (strain number: DLCS-31), whose growth condition and enzymatic properties were studied. The results showed that the optimum reaction pH and temperature of urate oxidase from strain DLCS-31 were 6.6 and 40 ℃, respectively. When incubated at 35 ℃ and 40 ℃ for 120 minutes, it maintained more than 80% of relative activity, and its activity increased to 140% when incubated at 40 ℃ for 60 minutes. The enzyme activity of strain DLCS-31 was about 4.3 times higher than that before optimization.