Cis/trans isomerisation of unsaturated fatty acids in a cardiolipin synthase knock-out mutant of Pseudomonas putida p8

• Published in: Applied microbiology and biotechnology Vol. 60; no. 1-2; pp. 179 - 185
• Main Authors: VON WALLBRUNN, A, HEIPIEPER, H. J, MEINHARDT, F
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01.10.2002; Springer Nature B.V
• Subjects: Adaptations; Bacterial genetics; Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; Biotechnology; Blotting, Southern; Chlorophenol; Cloning, Molecular; Electrophoresis, Polyacrylamide Gel; Fatty acids; Fatty Acids, Unsaturated - chemistry; Fatty Acids, Unsaturated - metabolism; Fundamental and applied biological sciences. Psychology; Genetic aspects; Genetics; High temperature; Isomerization; Lipids; Membrane Fluidity; Membrane Proteins; Microbiological synthesis; Mission oriented research; Mutants; Nutrition; Observations; Open Reading Frames; Organic solvents; Phenols; Pseudomonas putida; Pseudomonas putida - genetics; Pseudomonas putida - growth & development; Pseudomonas putida - metabolism; Sequence Analysis; Solvents; Stereoisomerism; Transferases (Other Substituted Phosphate Groups) - genetics; Transferases (Other Substituted Phosphate Groups) - metabolism; Germany; Pseudomonadales; Diphosphatidylglycerol; Cell proliferation; Acyltransferases; Enzyme; Transferases; Isomerization; Functional analysis; Fatty-acid synthase; Organic solvent; Production; Bacteria; Pseudomonadaceae; Unsaturated fatty acid; Molecular cloning; Pseudomonas putida; Sequencing
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-002-1080-y

The gene encoding cardiolipin synthase ( cls) from the phenol-degrading bacterium Pseudomonas putida P8, which rapidly adapts its membrane lipids to the presence of organic solvents by cis/trans isomerisation of unsaturated fatty acids, was isolated and completely sequenced. The functionality of the predicted gene product was proven by constructing a knock-out mutant that was significantly reduced in its growth rate both at elevated temperatures and in the presence of membrane-active solvents. Though the mutant showed a clear phenotype it was still able to synthesise trace amounts of cardiolipin. As an increase in cardiolipin (diphosphatidylglycerol) content is known to function as a long term membrane adaptation mechanism in pseudomonads, we tested whether the mutant compensates for the lack of the Cls by increased cis/trans isomerisation of unsaturated fatty acids. Increase in cis/trans isomerisation of unsaturated fatty acids was observed for the mutant at zero and low concentrations of 4-chlorophenol; however, cis/trans isomerisation is not able to fully compensate for the lack of cardiolipin production. Possibly, other long-term adaptation mechanisms are instrumental in compensating for the missing cardiolipin synthesis. As the cis/trans isomerase is activated similarly in the mutant and the wildtype, cis/trans isomerisation and cardiolipin production do not display mutual dependency.