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Degradation of diphenylether by Pseudomonas cepacia Et4: enzymatic release of phenol from 2,3-dihydroxydiphenylether

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Abstract

2,3-Dihydroxybiphenyl dioxygenase from Pseudomonas cepacia Et 4 was found to catalyze the ring fission of 2,3-dihydroxydiphenylether in the course of diphenylether degradation. The enzyme was purified and characterized. It had a molecular mass of 240 kDa and is dissociated by SDS into eight subunits of equal mass (31 kDa). The purified enzyme was found to be most active with 2,3-dihydroxybiphenyl as substrate and showed moderate activity with 2,3-dihydroxydiphenylether, catechol and some 3-substituted catechols. The K m-value of 1 μM for 2,3-dihydroxydiphenylether indicated a high affinity of the enzyme towards this substrate. The cleavage of 2,3-dihydroxydiphenylether by 2,3-dihydroxybiphenyl dioxygenase lead to the formation of phenol and 2-pyrone-6-carboxylate as products of ring fission and ether cleavage without participation of free intermediates. Isotope labeling experiments carried out with 18O2 and H2 18O indicated the incorporation of 18O from the atmosphere into the carboxyl residue as well as into the carbonyl oxygen of the lactone moiety of 2-pyrone-6-carboxylate. Based on these experimental findings the reaction mechanism for the formation of phenol and 2-pyrone-6-carboxylate is proposed in accordance with the mechanism suggested by Kersten et al. (1982).

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Abbreviations

DPE:

diphenylether

2,3-dihydroxy-DPE:

2,3-dihydroxydiphenylether

PCA:

2-pyrone-6-carboxylic acid

2,3-dihydroxy-BP dioxygenase:

2,3-dihydroxybiphenyl dioxygenase

GC:

gas chromatography

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Authors and Affiliations

  1. DMT-Gesellschaft für Forschung und Prüfung mbH, Institut für chemische Umwelttechnologie, Franz Fischer Weg 61, W-4300, Essen 13, Germany

    Frank Pfeifer, Jürgen Klein & Sigrid Schacht

  2. Institut für Mikrobiologie und Biotechnologie der Rheinischen Friedrich-Wilhelms-Universität, Meckenheimer Allee 168, W-5300, Bonn 1, Germany

    Hans G. Trüper

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  1. Frank Pfeifer
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  2. Hans G. Trüper
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  3. Jürgen Klein
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  4. Sigrid Schacht
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Pfeifer, F., Trüper, H.G., Klein, J. et al. Degradation of diphenylether by Pseudomonas cepacia Et4: enzymatic release of phenol from 2,3-dihydroxydiphenylether. Arch. Microbiol. 159, 323–329 (1993). https://doi.org/10.1007/BF00290914

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  • DOI: https://doi.org/10.1007/BF00290914

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