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Open AccessArticle

Metabolic Fingerprinting of Pseudomonas putida DOT-T1E Strains: Understanding the Influence of Divalent Cations in Adaptation Mechanisms Following Exposure to Toluene

Manchester Institute of Biotechnology, School of Chemistry, The University of Manchester, Manchester M1 7DN, UK
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Academic Editor: Peter Meikle
Metabolites 2016, 6(2), 14; https://doi.org/10.3390/metabo6020014
Received: 16 February 2016 / Revised: 20 April 2016 / Accepted: 21 April 2016 / Published: 26 April 2016
(This article belongs to the Special Issue Cancer Metabolomics 2016)
Pseudomonas putida strains can adapt and overcome the activity of toxic organic solvents by the employment of several resistant mechanisms including efflux pumps and modification to lipopolysaccharides (LPS) in their membranes. Divalent cations such as magnesium and calcium play a crucial role in the development of solvent tolerance in bacterial cells. Here, we have used Fourier transform infrared (FT-IR) spectroscopy directly on cells (metabolic fingerprinting) to monitor bacterial response to the absence and presence of toluene, along with the influence of divalent cations present in the growth media. Multivariate analysis of the data using principal component-discriminant function analysis (PC-DFA) showed trends in scores plots, illustrating phenotypic alterations related to the effect of Mg2+, Ca2+ and toluene on cultures. Inspection of PC-DFA loadings plots revealed that several IR spectral regions including lipids, proteins and polysaccharides contribute to the separation in PC-DFA space, thereby indicating large phenotypic response to toluene and these cations. Finally, the saturated fatty acid ratio from the FT-IR spectra showed that upon toluene exposure, the saturated fatty acid ratio was reduced, while it increased in the presence of divalent cations. This study clearly demonstrates that the combination of metabolic fingerprinting with appropriate chemometric analysis can result in practicable knowledge on the responses of important environmental bacteria to external stress from pollutants such as highly toxic organic solvents, and indicates that these changes are manifest in the bacterial cell membrane. Finally, we demonstrate that divalent cations improve solvent tolerance in P. putida DOT‑T1E strains. View Full-Text
Keywords: fingerprinting; efflux pumps; P. putida DOT-T1E; toluene; stress tolerance; LPS; Mg2+; Ca2+; FT-IR fingerprinting; efflux pumps; P. putida DOT-T1E; toluene; stress tolerance; LPS; Mg2+; Ca2+; FT-IR
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MDPI and ACS Style

Sayqal, A.; Xu, Y.; Trivedi, D.K.; AlMasoud, N.; Ellis, D.I.; Goodacre, R. Metabolic Fingerprinting of Pseudomonas putida DOT-T1E Strains: Understanding the Influence of Divalent Cations in Adaptation Mechanisms Following Exposure to Toluene. Metabolites 2016, 6, 14.

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