Process for the flooding of petroleum deposits and oil shale by means of dispersions of non-ionogenic, boundary surface active materials in water as flooding medium, through the utilization of a suitable bioreactor with or without mechanical stirring for a first stage for the production of a growing submerged culture of hydrocarbon enriched microorganisms in a semi- or continuous process under aerobic conditions, under the addition of organic C- sources, inorganic nutrients dissolved in water and occasionally growth materials as well as the addition of air or oxygen-enriched air at a predetermined reaction temperature and constant pH value in the region of between 2 and 9, with a second stage in which the formed effective material is separated from the cell material and, with utilization of the aqueous separating phase with the therein dispersed effective material directly as flooding medium or as addition to the flooding water, characterized in that in the first stage there are produced as the effective material glycolipids with the hydrocarbon enriched microorganism under the utilization of a hydrocarbon mixture of 1 to 35% by volume at a reaction temperature in the range of 20.degree. to 50.degree. C. and at a constant pH value in the range of between 3 and 9, and that thereafter in the second stage the formed glycolipids are separated from the cell material in the form of an aqueous phase through a temperature, pH, or osmotic shock or through extraction and a petroleum deposit and oil shale flooding amount of the aqueous phase with the therein dispersed glycolipids are directly used as a flooding medium or added to the flooding water.