Fatty acid biosynthesis is involved in solvent tolerance in Pseudomonas putida DOT-T1E

Ana Segura, Estrella Duque, Antonia Rojas, Patricia Godoy, Antonio Delgado, Ana Hurtado, John E. Cronan, Juan Luis Ramos

Research output: Contribution to journalArticlepeer-review

Abstract

The unusual tolerance of Pseudomonas putida DOT-T1E to toluene is based on the extrusion of this solvent by constitutive and inducible efflux pumps and rigidification of its membranes via phospholipid alterations. Pseudomonas putida DOT-T1E-109 is a solvent-sensitive mutant. Mutant cells were less efficient in solvent extrusion than the wild-type cells, as shown by the limited efflux of 14C-1,2,4-trichlorobenzene from the cell membranes, despite the fact that the efflux pumps are overexpressed as a result of increased expression of the ttgDEF and ttgGHI efflux pump operons. This limitation could be the result of alterations in the outer membrane because the mutant cells released more β-lactamase to the external medium than the wild-type cells.The mutant P. putida DOT-T1E-109 showed negligible synthesis of fatty acids in the presence of sublethal concentrations of toluene as revealed by analysis of 13CH3-13COOH incorporation into fatty acids. In contrast, the mutant strain in the absence of solvents, and the wild-type strain, both in the presence and in the absence of toluene, incorporated 13CH3-13COOH at a high rate into de novo synthesized lipids.The mutation in P. putida DOT-T1E-109 increases sensitivity to the solvent because of a limited efflux of the solvent from the cell membranes with the concomitant inhibition of fatty acid biosynthesis.

Original languageEnglish (US)
Pages (from-to)416-423
Number of pages8
JournalEnvironmental Microbiology
Volume6
Issue number4
DOIs
StatePublished - Apr 1 2004

ASJC Scopus subject areas

  • Microbiology
  • Ecology, Evolution, Behavior and Systematics

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