Bacterial bio-surfactants have a wide range of biological functions and biotechnological applications. Previous analyses had suggested a limit to their reduction of aqueous liquid surface tensions (gMin), and here we confirm this in an analysis of 25 Pseudomonas spp. strains isolated from soil which produce high-strength surfactants that reduce surface tensions to 25.2 ± 0.1 – 26.5 ± 0.2 mN.m-1 (the surface tension of sterile growth medium and pure water was 52.9 ± 0.4 mN.m-1 and 72.1 ± 1.2 mN.m-1, respectively). Comparisons of culture supernatants produced using different growth media and semi-purified samples indicate that the limit of 24.2 – 24.7 mN.m-1 is not greatly influenced by culture conditions, pH or NaCl concentrations. We have used foam, emulsion and oil-displacement behavioural assays as a simple and cost-effective proxy for in-depth biochemical characterisation, and these suggest there is significant structural diversity amongst these surfactants which may reflect different biological functions and offer new biotechnological opportunities. Finally, we obtained a draft genome for the strain producing the highest-strength surfactant, and identified a cluster of non-ribosomal protein synthase genes which may produce a cyclic-lipopeptide (CLP)–like surfactant. Further investigation of this group of related bacteria recovered from the same site will allow a better understanding of the significance of the great variety of surfactants produced by bacterial communities found in soil and elsewhere.