Rhamnolipids are produced by the bacterium Pseudomonas aeruginosa and several other bacteria. Since P. aeruginosa is found naturally in nature in soils, water and on plants, the presence of rhamnolipids is also common although in very small quantities.

It has been discovered that several other species of bacteria can produce rhamnolipids under the correct conditions. These species include Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas oleovorans,Pseudomonas chlororaphis, Renibacterium salmoninarum, and Burkholderia pseudomallei (previously known as Pseudomonas pseudomallei.), and others

All of these species, except P. chlororaphis, are considered pathogenic to human and animal bodies. P. putida is has been found in association with certain cancers and is therefore considered a carcinogenic pathogen.

These all are capable of extra cellular secretion of rhamnolipids, when growing on soluble and insoluble carbon sources.

Although, P. chlororaphis, which is not considered a pathogen, can produce rhamnolipids, it can only produce the rhamnolipids in very low quantities. It cannot produce di-rhamnolipids.

Because P. aeruginosa can be used to produce rhamnolipids in the highest quantity, it is the species used for rhamnolipid production. The other advantage of P. aeruginosa is that it produces more types of both mono-rhamnolipids and di-rhamnolipids that any other bacterium.

There has been work transplanting the genes responsible for controlling the production of rhamnolipids in to other bacteria and plants. This is considered genetic modification (GM). The use of the resultant organism or material is banned or regulated by many government authorities such as the European Union. Rhamnolipid, Inc. does NOT use or supply any such GM organisms or material.

Because bacterium P. aeruginosa itself is a pathogen and can make people and animals sick in sufficient quantities, the resulting rhamnolipids are sterilized and separated from the bacteria so that the product is safe.