Bacterial preoxidation of refractory gold ore has showed many advantages, such as lower operation cost and environmental friendly. In this paper, biooxidation of refractory gold ore by mixed moderate thermophiles culture in airlift bioreactor has been well studied. Firstly, a laboratory scale airlift bioreactor unit has been designed for bacterial preoxidation of refractory gold ore, a mixed moderate thermophilic culture was incubated. The orthogonal experiment was performed to investigate the effects of particle size, pulp density, aeration rate and initial pH. The results showed that when particle size 37micrometer，pulp density 5%，aeration rate 4 dm3•s-1 and initial pH 1.2, the arsenic leaching rate achieved to 93.5%. On the base of the airlift bioreactor unit, a continuous airlift bioreactor was designed. The preoxidation of refractory gold ore by a mixed culture of moderately thermophilic bacteria also was investigated in this equipment. The results showed that, under the conditions of pulp density 8% and stay time 60h, it can deal with 0.256 kg refractory gold ore per day, and the arsenic leaching rate can reach to 95.25%. During the experiment, some leaching liquor has been sampled from the second sect the fourth column of the continuous airlift bioreactor. The composition of microbial communities in this sample has been investigated using culture-independent 16S rDNA based cloning approach- restriction fragment length polymorphism. The phylogenetic analysis revealed that Leptospirillum ferriphilum and Acidithiobacillus caldus are the main organisms in this system. Both of them are moderate thermphilic bacteria and can resist relatively high concentration of arsenic ion.