We review the environmental role of rhamnolipids in terms of microbial life and activity. A large number of previous research supports the idea that these glycolipids mediate the uptake of hydrophobic substrates by bacterial cells. This feature might be of highest priority for bioremediation of spilled hydrocarbons. However, current evidence confirms that rhamnolipids primarily play a role in surface-associated modes of bacterial motility and are involved in biofilm development. This might be an explanation why no direct pattern of hydrocarbon degradation was often observed after rhamnolipids supplementation. This review gives insight into the current state of knowledge on how rhamnolipids operate in the microbial world.