One Bacillus strain named SLY-3 producing bacteriocin-like substance (BLS) was screened from the soil polluted by oil. The G bacteria and molds were effectively inhibited by BLS produced by strain SLY-3. With the analysis of colony morphology, physiological and biochemistry experiments and 16S rDNA gene sequence, the strain SLY-3 was identified as Bacillus subtilis. The BLS secreted by Bacillus subtilis SLY-3 had the highest inhibitory activity when it was cultured for 24 h at initial pH 7.0 and 28℃.