Pseudomonas aeruginosa presents three types of motilities: swimming, twitching and swarming. The latter is characterized by rapid and coordinated group movement over a semisolid surface resulting from morphological differentiation and intercellular interactions. A striking feature of P. aeruginosa swarming motility is the formation of migrating tendrils producing colonies with complex fractal-like patterns. Previous studies have shown that normal swarming motility is intimately related to the production of extracellular surface-active molecules: rhamnolipids (RLs), composed of monorhamnolipids (mono-RLs) and dirhamnolipids (di-RLs), and 3-(3-hydroxyalkanoyloxy) alkanoic acids (HAAs). Here, we report that (i) di-RLs attract active swarming cells while HAAs behave as strong repellents, (ii) di-RLs promote and HAAs inhibit tendril formation and migration, (iii) di-RLs and HAAs display different diffusion kinetics on a surface as di-RLs spread faster than HAAs in agar, (iv) di-RLs and HAAs have no effect on swimming cells, suggesting that swarming cells are different from swimming cells not only in morphology but also at the regulatory level and (v) mono-RLs act as wetting agents. We propose a model explaining how HAAs and di-RLs together modulate the behaviour of swarming migrating cells by acting as self-produced negative and positive chemotactic-like stimuli.

Pseudomonas aeruginosa produces the biosurfactants rhamnolipids and 3-(3-hydroxyalkanoyloxy)alkanoic acids (HAAs). In this study, we report the production of one family of rhamnolipids, specifically the monorhamnolipids, and of HAAs in a recombinant Escherichia coli strain expressing P. aeruginosa rhlAB operon. We found that the availability in E. coli of dTDP-L: -rhamnose, a substrate of RhlB, restricts the production of monorhamnolipids in E. coli. We present evidence showing that HAAs and the fatty acid dimer moiety of rhamnolipids are the product of RhlA enzymatic activity. Furthermore, we found that in the recombinant E. coli, these compounds have the same chain length of the fatty acid dimer moiety as those produced by P. aeruginosa. These data suggest that it is RhlAB specificity, and not the hydroxyfatty acid relative abundance in the bacterium, that determines the profile of the fatty acid moiety of rhamnolipids and HAAs. The rhamnolipids level produced in recombinant E. coli expressing rhlAB is lower than the P. aeruginosa level and much higher than those reported by others in E. coli, showing that this metabolic engineering strategy lead to an increased rhamnolipids production in this heterologous host.