A field investigation was performed to study the content, speciation and mobility of vanadium, as well as microbial response in soil from a stone coal smelting district in Hunan province of China. The results showed that the contents of soil V ranged from 168 to 1538 mg/kg, which exceeded the maximum permissible value of Canadian soil quality for V. The mean soil V content from wasteland area reached 1421 mg/kg, and those from the areas related with slag heap, ore pile and smelting center were 380, 260 and 225 mg/kg, respectively. Based on the results of the modified BCR sequential extraction procedure, V contents in the mobile fractions varied from 19.2 to 637 mg/kg accounting for 7.4%-42.3% of total V, and those of V(+5) species were between 21.9 and 534.0 mg/kg. Soil enzyme activity and microbial basal respiration were adversely affected by high level of soil V. More attention should be paid to soil V pollution and potential hazardous surrounding the stone coal smelting district.