Pseudomonas putida strains are prevalent in a variety of pristine and polluted environments. The genome of the solvent‐tolerant P. putida strain DOT‐T1E which thrives in the presence of high concentrations of monoaromatic hydrocarbons, contains a circular 6.3 Mbp chromosome and a 133 kbp plasmid. Omics information has been used to identify the genes and proteins involved in solvent tolerance in this bacterium. This strain uses a multifactorial response that involves fine‐tuning of lipid fluidity, activation of a general stress‐response system, enhanced energy generation, and induction of specific efflux pumps that extrude solvents to the medium. Local and global transcriptional regulators participate in a complex network of metabolic functions, acting as the decision makers in the response to solvents.

In Pseudomonas putida, as in many other eubacteria, cyclopropane fatty acids (CFAs) accumulate in the membrane during the stationary phase of growth. Here, we show that cfaB gene expression in P. putida KT2440 is dependent on the RpoS sigma factor that recognizes the sequence 5'-CTACTCT-3' between -8 and -14. We have carried out a mutational study of the cfa promoter and have determined that positions -9, -12, -13 and -14 are the most critical for maximal activity. In P. putida, the substrates of the CFA synthase, cis-unsaturated fatty acids (cis-UFAs), are also substrates for another stress-related enzyme, the cis-trans isomerase (CTI). Despite using the same substrates, we have found that the activity of the CTI is not limited by the CFA synthase activity and vice versa. For instance, in a cfaB knockout mutant, the amount of trans-UFAs synthesized after a specific stress was no higher than in the parental background despite the fact that there are more cis-UFAs available to be used by the CTI as substrates. In this regard, in a cti-deficient mutant background, the levels of CFAs were similar to those in the parental one under the same conditions.

Bacteria of the species Pseudomonas putida are ubiquitous soil inhabitants, and a few strains are able to thrive in the presence of extremely high concentrations of toxic solvents such as toluene and related aromatic hydrocarbons. Toluene tolerance is multifactorial in the sense that bacteria use a wide range of physiological and genetic changes to overcome solvent damage. This includes enhanced membrane impermeabilization through cis to trans isomerization of unsaturated fatty acids, activation of a stress response programme, and induction of efflux pumps that expulse toxic hydrocarbons to the outer medium. The most relevant element in this toluene tolerance arsenal is the TtgGHI efflux pump controlled by the TtgV regulator. We discuss here how TtgV controls expression of this efflux pump in response to solvents.

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