Several hundred bacterial cultures isolated from a variety of natural sites were screened for their ability to desulfurize the model coal organosulfur compounds, dibenzothiophene (DBT) and DBT-sulfone. A sulfur-stress assay, in which DBT-sulfone was the only bioavailable source of sulfur, was used to screen and select for organisms that selectively desulfurized the organic-sulfur substrate. Only two new isolates, UMX9 and UMX3, and strain IGTS-8, a Rhodococcus rhodochrous provided by the Institute for Gas Technology (Chicago, USA.) as a reference culture, would grow on DBT or DBT-sulfone as a sole source of sulfur. Under sulfur-stress conditions, a desulfurized product identified as 2-hydroxybiphenyl (2-phenylphenol) was detected only for UMX9 and IGTS-8. Biodesulfurization activity for all three organisms occurred only for growing cultures, and was depressed by free sulfate, although more so for UMX3 and IGTS-8 than for UMX9. None of the three cultures exhibited good growth on DBT, DBT-sulfone, or 2-phenylphenol as sole sources of carbon. Taxonomic studies revealed UMX3 to be similar to IGTS-8, whereas UMX9 only exhibited Rhodococcus-like features. Comparative tests for carbohydrate utilization revealed that only UMX9 would grow on glucose, and that only IGTS-8 would grow on L-arabinose. Assays of biodesulfurization activity as a function of temperature ormore » pH revealed further differences between UMX9 and UMX3/IGTS-8. Under optimized assay conditions for each organism, UMX9 exhibited up to 30% greater biodesulfurization activity than did IGTS-8 and UMX3, which were similar in activity.« less