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The petrochemical industry is essential for many consumer goods currently in use. To implement innovative biobased processes, renewable resources are used as substitutes for crude oil in goods such as cleaning agents. However, biosurfactants are still expensive compared to their chemically synthesized counterparts. In this work, the exploitation of Pseudomonas putida for the heterologous synthesis of rhamnolipids, glycolipids with a high potential for industrial applications was the main focus.