The bioleaching of a high alkaline copper oxide ore from Yunnan Province in southwest China was carried out in a shake flask with ammonia-producing bacteria Provideneia JAT-1. It is found that temperature, liquid-solid ratio, species and concentration of the leaching aid, and initial bacteria inoculation have significant impact on the copper leaching rate. When the temperature is 30℃, the liquid-solid mass ratio is 7:1, the concentration of ammonium sulfate as the leaching aid is 0.024 mol·L^-1, and the initial bacteria inoculation is 20%, the bioleaching for 144 h yields a copper recovery of 42.35%. The result of phase analysis shows that the copper leaching rate of secondary sulphide copper in the ore is the highest.