Fatty acid composition in Pseudomonas sp. CF600 during degradation of catechol and phenol individually and their mixture was investigated. Moreover, the influence of glucose as an additional, easily degradable carbon source on fatty acid profiling in bacteria grown on these aromatic substrates was studied. Both catechol and phenol treatments caused in bacterial cells crucial changes in the distribution of tested groups of fatty acids. The major changes included the increase of fatty acid saturation, decrease in the percentage of cyclopropane fatty acid 17:0 cy and the appearance of branched and hydroxy fatty acids. Under catechol, phenol and their mixture exposure saturated/unsaturated ratio showed the value 6.5, 5.68 and 6.38 whereas in control cells this ratio reached the value 3.05. As a response to aromatic compounds bacteria formed fatty acids that were not detected in control cells growing on glucose. It has been demonstrated that the supplementation of cultured media containing single aromatic substrates or/and their mixture with glucose resulted in changes in degradation rates of catechol and phenol. It seemed that glucose influenced some metabolic pathways responsible for the assimilation of aromatic compounds. The incubation of cells in the presence of aromatic compounds and glucose rapidly led to alterations of whole-cell derived fatty acid composition. The most important changes were associated with saturation level of fatty acids and cyclopropane fatty acid contents.