The microbial biosurfactants can be substituted to the chemical detergents in some industrial processes. In this study we developed a biotechnological processes for the biosurfactants with microorganisms. The biosurfactants have a lot of advantages in comparision with the chemical surfactants. They are proenvironmental even during and after industrial use. But there are not so many kinds of biosurfactants. The production cost and the end price is much higher than the chemical surfactants. But nowdays there are many kinds of microorganisms, which can produce the surfactants in large quantity and fast. We tried to develop a production process for the large scale with some microorganisms. At first Candida bombicola KCTC 7145, Sphingomonas chungbukensis KCTC 2955 and Sphingomonas yanoikuyae KCTC 2818 are cultivated and studied. For the large scale production process we used molasses as a complex medium and tried to optimize the process. Molasses contains 17 to 25% of water, 45 to 50% of sugar and 25% of carbohydrate, it can be fully used as a substrate. The microorganisms have been cultivated in the diluted media with molasses 2, 5, 8 and 10%, respectively, The optimal conditions for the cultivation and the production process have been studied. For the study the optical density, glucose concentration and the surface tension were measured. Candida bombicola KCTC 7145 and the 5% molasses media was selected as an optimal condition for the production process of a biosurfactant. During cultivation of Candida bombicola KCTC 7145 in the 5% molasses medium kerosene and corn oil were added for promoting the biosurfactants.