New developments in biotechnology are focused on new products or the introduction of new processes as a substitute to the conventional way, or improvements in performance of existing biotechnological applications. The examples of application of non-conventional media are organic solvents, supercritical solvents or aqueous two phases systems. Steroid biotransformation is a multi million-dollar industry and the pharmaceutical uses of steroids are numerous. Steroids like cholesterol are completely soluble in some organic solvents like benzene, toluene and butanol. Biphasic systems wherein the cells are present in the aqueous phase and steroids dissolved in the organic phase is considered an ideal set-up. The major problem here is the fact that most bacteria and their enzymes are inactivated or destroyed in the presence of these toxic organic solvents, also it is necessary that the microbes contain relevant enzymes in sufficient amounts and in high specific activities, even in the presence of the generally destructive apolar phase. This can be very well accomplished by using organic solvent tolerant bacteria having desired enzyme activities. One such application is being discussed.