Biosurfactants are of considerable industrial value as their high tenside activity in combination with their biocompatibility makes them attractive for many applications. In particular members of the lipopeptide family of biosurfactants contain significant potentials for the pharmaceutical industry due to their intrinsic antibiotic characteristics. The high frequency of lipopeptide (LP) production in common soil microorganisms in combination with the enormous structural diversity of the synthesized biosurfactants has created an abundant natural pool of compounds with potentially interesting properties. Unfortunately, the bioactivity of lipopetides against pathogenic microorganisms is often associated with problematic side effects that restrict or even prevent medically relevant applications. The accumulated knowledge of lipopetide biosynthesis and their frequent structural variations caused by natural genetic rearrangements has therefore motivated numerous approaches in order to manipulate biosurfactant composition and production mechanisms. This chapter will give an overview on current engineering strategies that aim to obtain lipopeptide biosurfactants with redesigned structures and optimized properties.