It is very important that selection of packing media with large surface area, high limited back pressure in biofilter. The object of this study is the isolation of sulfur-oxidizing bacteria and the removal of hydrogen sulfide in biofilter by media. This investigation led to the following results: 1) we isolated Thiobacillus sp. IW. at an abandoned coal mine in Hwasun, Jeonnam Province. 2) The inorganic media showed better results than the organic media from experiments looking at removal characteristics and changes in pressure drop using various media. 3) Among the inorganic media, fibril and PU media showed best performance.