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NMR spectroscopy as a tool to investigate the degradation of aromatic compounds by a Pseudomonas putida strain
Author(s) -
Bertini I.,
Provenzani A.,
Viezzoli M. S.,
Pieper D. H.,
Timmis K. N.
Publication year - 2003
Publication title -
magnetic resonance in chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.483
H-Index - 72
eISSN - 1097-458X
pISSN - 0749-1581
DOI - 10.1002/mrc.1216
Subject(s) - pseudomonas putida , chemistry , strain (injury) , nuclear magnetic resonance spectroscopy , spectroscopy , pseudomonas , pseudomonadaceae , two dimensional nuclear magnetic resonance spectroscopy , carbon 13 nmr , bacteria , nuclear chemistry , organic chemistry , chromatography , stereochemistry , medicine , physics , quantum mechanics , enzyme , biology , genetics
Pseudomonas putida strain KT2442, harbouring the pWW0 TOL plasmid, was grown with a number of different homologous aromatic acids as carbon sources. Small samples of liquid culture supernatant were collected and directly analysed by 2D NMR spectroscopy. In all cases similar compounds with olefinic signals were observed to accumulate. To elucidate the structures of these compounds, 2D NMR experiments with 500 and 600 MHz spectrometers equipped with a CryoProbe (Bruker BioSpin) were performed on samples obtained from a culture growing on 4‐methylbenzoate and, for 13 C spectroscopy, on 13 C‐labelled 4‐methylbenzoate. In all cases a 1,2‐dihydroxycyclohexa‐3,5‐diene‐carboxylate derivative was identified. The use of this technique helped us to identify easily some metabolites that were released into the solution by bacteria and to follow their secretion as a function of time. The high sensitivity of the present approach allowed a clear and rapid acquisition of spectra, notwithstanding the low concentration of the compounds. The benefits of introducing the use of NMR cryoprobes to perform metabolic pathway studies is demonstrated. Copyright © 2003 John Wiley & Sons, Ltd.