Rhamnolipids are amphiphilic surface active molecules that are currently receiving major interest from industry as a sustainable alternative to chemical surfactants produced using petrochemical feedstock's. Whilst the majority of research has been carried out on P. aeruginosa in terms of industrial scale RL production the organisms' pathogenic nature makes its use on an industrial scale uneconomic. The main focus of this thesis was therefore to explore whether B, thailandensis, a non-pathogenic alternative to P. aeruginosa, could be characterized and manipulated in order to produced RLs at a level equal to or greater than that of P. aeruginosa. The first part of this thesis was used to characterize RL production in B. thailandensis and to gauge its potential as a possible industrial scale RL producer. As very little work has been carried out on B. thailandensis in terms of RL production this chapter was used to generate a detailed, quantitative outlineof RL production in B. thailandensis under specific growth conditions as well as giving an insight into some of the phenotypic traits RLs are involved in. In addition, quantitative expression analysis of RL synthesis genes also provides a transcriptional profile of RL production providing a complete and in depth profile of RL biosynthesis in B. thailandensis E264.