ABSTARCT: Microbially produced lipopeptide have been isolated and studied for microbial enhanced oil recovery.About 60 gram positive bacteria isolated from soil contaminated with crude oil, near the crude oil storage tank in Tehran Refinery, Tehran, Iran. However, most of these studies have produced lipopeptide by one of the pure-culture microbes isolated in a laboratory. Among the isolates, heamolytic tests revealed two biosurfactant producers. The isolated strains were designated as C2, E1. By using morphological, biochemical and molecular biology tests (16 SrRNA), the strains identified as Bacillus licheniformis and Bacillus subtitlis, respectively. Emulsification activity and measurement of surface tension indicated that, the isolates were high producers of biosurfactant. The product of C2 and E1 is mainly lipopeptide. This product reduce surface tension from 65 to 30 mN/m. Emulsified activity of crude oil was 92% for C2 and 90 % in case of E1. This is the first report of indigenous Bacillus licheniformis and Bacillus subtilis from a soil contaminated with oil in an Iranian refinery with ability to produce biosurfactant.