AbstractBiocatalytic Desulfurization (BDS) represents an alternative approach to the reduction of sulfur in fossil fuels. The objective is to use bacteria to selectively remove sulfur from petroleum and middle distillate fractions, without the concomitant release of carbon. Recently, bacteria have been developed which have the ability to desulfurize dibenzothiophene (DBT) and other organosulfur molecules. These bacteria are being developed for use in a biocatalyst‐based desulfurization process. Analysis of preliminary conceptual engineering designs has shown that this process has the potential to complement conventional technology as a method to temper the sulfur levels in crude oil, or remove the recalcitrant sulfur in middle distillates to achieve the deep desulfurization mandated by State and Federal regulations.This paper describes the results of initial feasibility studies, sensitivity analyses and conceptual design work. Feasibility studies with various crude oils and middle distillates achieved unoptimized desulfurization levels of 40–80%. Sensitivity analyses indicate that total desulfurization costs of about $3.00 per barrel for crude oil and less than $2.00 per barrel for diesel are possible. Key criteria for commercial success of the process include the cost and half‐life of the biocatalyst, residence time in the reactor, oil/water ratios required to extract the sulfur and the disposition of the separated sulfur pro