Chemical surfactants are ubiquitous in consumer products, but they are the subject of environmental concerns. For this reason, the complete replacement of petrochemical surfactants with biosurfactants is a holy grail, but this is far from possible in the near future. Although the “biosurfactant revolution” has not yet taken place, mainly due to the higher cost and lower availability of biosurfactants, another reason could explain this fact: poor knowledge of their properties in solution. The aim of this tutorial is to test the self-organization properties and phase behavior experimentally (Sections 3.3 and 3.4) and through molecular modeling (Section 6) of the most important microbial biosurfactants (sophorolipids, rhamnolipids, surfactin, cellobiose lipids, glucolipids) in water) and their Main derivatives. A critical discussion of such properties in the light of the known packing parameters of surfactants is also given (Section 3.5). The relationship between nanoscale self-organization and the properties of the macroscopic material, including hydrogelling, rigid foaming, templating, or encapsulation, is specifically discussed (Section 3.7). We present their self-organization and adsorption even with flat and complex air / liquid (e.g., Foams), air / solid (adhesion), liquid / solid (nanoparticles) and liquid / liquid (e.g., Emulsions) interfaces (Section 4). A critical discussion about the use of biosurfactants as capping agents for the development of stable nanoparticles is given specifically (Section 4.2.4). Finally, we discuss the key findings from biosurfactants and macromolecules, including proteins, enzymes, polymers, and polyelectrolytes.