Comparative analysis of rhamnolipids from novel environmental isolates of Pseudomonas aeruginosa

A comparative analysis of rhamnolipids from environmental isolates of Pseudomonas aeruginosa was undertaken to evaluate strain-specific rhamnolipid fingerprints obtained under different growth conditions. Environmental isolates of P. aeruginosa produced rhamnolipids on different types of substrates, including cheap and renewable sources like sunflower oil from deep fryers and sunflower oil mill effluent. Rhamnolipids were monitored by high-performance liquid chromatography–electrospray ionization interface mass spectrometry, which allowed fast and reliable identification and quantification of the congeners present. The highest concentration of total rhamnolipids of 3.33 g/l was obtained by the strain P. aeruginosa 67, recovered from petroleum contaminated soil, and strains D1 (1.73 g/l) and D2 (1.70 g/l), recovered from natural microbial consortia originated from mazut-contaminated soil, grown on sunflower oil as a carbon source. Di- to mono-rhamnolipids ratios were in the range of 0.90–5.39 for different media composition and from 1.12 to 4.17 for different producing strains. Rhamnolipid profiles of purified mixtures of all tested strains are similar with chain length from C8–C12, pronounced abundance of Rha–C10–C10 and Rha–Rha–C10–C10 congeners, and a low content of 3-(3-hydroxyalkanoyloxy)-alkanoic acids. Concentrations of major congeners of RLs were found to slightly vary, depending on strain and growth conditions, while variations in minor congeners were more pronounced. Statistically significant increase of critical micelle concentration values was observed with lowering the ratio of total mono- to di-rhamnolipids ratio indicating that mono-rhamnolipids start to form micelles at lower concentration than di-rhamnolipids.