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Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium Thauera aromatica
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| Zeitschriftentitel: | Journal of Bacteriology |
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| Personen und Körperschaften: | , , |
| In: | Journal of Bacteriology, 182, 2000, 20, S. 5849-5863 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
American Society for Microbiology
|
| Schlagwörter: |
| author_facet |
Breinig, Sabine Schiltz, Emile Fuchs, Georg Breinig, Sabine Schiltz, Emile Fuchs, Georg |
|---|---|
| author |
Breinig, Sabine Schiltz, Emile Fuchs, Georg |
| spellingShingle |
Breinig, Sabine Schiltz, Emile Fuchs, Georg Journal of Bacteriology Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium Thauera aromatica Molecular Biology Microbiology |
| author_sort |
breinig, sabine |
| spelling |
Breinig, Sabine Schiltz, Emile Fuchs, Georg 0021-9193 1098-5530 American Society for Microbiology Molecular Biology Microbiology http://dx.doi.org/10.1128/jb.182.20.5849-5863.2000 <jats:title>ABSTRACT</jats:title> <jats:p> Genes involved in the anaerobic metabolism of phenol in the denitrifying bacterium <jats:italic>Thauera aromatica</jats:italic> have been studied. The first two committed steps in this metabolism appear to be phosphorylation of phenol to phenylphosphate by an unknown phosphoryl donor (“phenylphosphate synthase”) and subsequent carboxylation of phenylphosphate to 4-hydroxybenzoate under release of phosphate (“phenylphosphate carboxylase”). Both enzyme activities are strictly phenol induced. Two-dimensional gel electrophoresis allowed identification of several phenol-induced proteins. Based on N-terminal and internal amino acid sequences of such proteins, degenerate oligonucleotides were designed to identify the corresponding genes. A chromosomal DNA segment of about 14 kbp was sequenced which contained 10 genes transcribed in the same direction. These are organized in two adjacent gene clusters and include the genes coding for five identified phenol-induced proteins. Comparison with sequences in the databases revealed the following similarities: the gene products of two open reading frames (ORFs) are each similar to either the central part and N-terminal part of phosphoenolpyruvate synthases. We propose that these ORFs are components of the phenylphosphate synthase system. Three ORFs showed similarity to the <jats:italic>ubiD</jats:italic> gene product, 3-octaprenyl-4-hydroxybenzoate carboxy lyase; UbiD catalyzes the decarboxylation of a 4-hydroxybenzoate analogue in ubiquinone biosynthesis. Another ORF was similar to the <jats:italic>ubiX</jats:italic> gene product, an isoenzyme of UbiD. We propose that (some of) these four proteins are involved in the carboxylation of phenylphosphate. A 700-bp PCR product derived from one of these ORFs cross-hybridized with DNA from different <jats:italic>Thauera</jats:italic> and <jats:italic>Azoarcus</jats:italic> strains, even from those which have not been reported to grow with phenol. One ORF showed similarity to the <jats:italic>mutT</jats:italic> gene product, and three ORFs showed no strong similarities to sequences in the databases. Upstream of the first gene cluster, an ORF which is transcribed in the opposite direction codes for a protein highly similar to the DmpR regulatory protein of <jats:italic>Pseudomonas putida</jats:italic> . DmpR controls transcription of the genes of aerobic phenol metabolism, suggesting a similar regulation of anaerobic phenol metabolism by the putative regulator. </jats:p> Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium <i>Thauera aromatica</i> Journal of Bacteriology |
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American Society for Microbiology, 2000 |
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American Society for Microbiology, 2000 |
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| title |
Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium Thauera aromatica |
| title_unstemmed |
Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium Thauera aromatica |
| title_full |
Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium Thauera aromatica |
| title_fullStr |
Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium Thauera aromatica |
| title_full_unstemmed |
Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium Thauera aromatica |
| title_short |
Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium Thauera aromatica |
| title_sort |
genes involved in anaerobic metabolism of phenol in the bacterium
<i>thauera aromatica</i> |
| topic |
Molecular Biology Microbiology |
| url |
http://dx.doi.org/10.1128/jb.182.20.5849-5863.2000 |
| publishDate |
2000 |
| physical |
5849-5863 |
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<jats:title>ABSTRACT</jats:title>
<jats:p>
Genes involved in the anaerobic metabolism of phenol in the denitrifying bacterium
<jats:italic>Thauera aromatica</jats:italic>
have been studied. The first two committed steps in this metabolism appear to be phosphorylation of phenol to phenylphosphate by an unknown phosphoryl donor (“phenylphosphate synthase”) and subsequent carboxylation of phenylphosphate to 4-hydroxybenzoate under release of phosphate (“phenylphosphate carboxylase”). Both enzyme activities are strictly phenol induced. Two-dimensional gel electrophoresis allowed identification of several phenol-induced proteins. Based on N-terminal and internal amino acid sequences of such proteins, degenerate oligonucleotides were designed to identify the corresponding genes. A chromosomal DNA segment of about 14 kbp was sequenced which contained 10 genes transcribed in the same direction. These are organized in two adjacent gene clusters and include the genes coding for five identified phenol-induced proteins. Comparison with sequences in the databases revealed the following similarities: the gene products of two open reading frames (ORFs) are each similar to either the central part and N-terminal part of phosphoenolpyruvate synthases. We propose that these ORFs are components of the phenylphosphate synthase system. Three ORFs showed similarity to the
<jats:italic>ubiD</jats:italic>
gene product, 3-octaprenyl-4-hydroxybenzoate carboxy lyase; UbiD catalyzes the decarboxylation of a 4-hydroxybenzoate analogue in ubiquinone biosynthesis. Another ORF was similar to the
<jats:italic>ubiX</jats:italic>
gene product, an isoenzyme of UbiD. We propose that (some of) these four proteins are involved in the carboxylation of phenylphosphate. A 700-bp PCR product derived from one of these ORFs cross-hybridized with DNA from different
<jats:italic>Thauera</jats:italic>
and
<jats:italic>Azoarcus</jats:italic>
strains, even from those which have not been reported to grow with phenol. One ORF showed similarity to the
<jats:italic>mutT</jats:italic>
gene product, and three ORFs showed no strong similarities to sequences in the databases. Upstream of the first gene cluster, an ORF which is transcribed in the opposite direction codes for a protein highly similar to the DmpR regulatory protein of
<jats:italic>Pseudomonas putida</jats:italic>
. DmpR controls transcription of the genes of aerobic phenol metabolism, suggesting a similar regulation of anaerobic phenol metabolism by the putative regulator.
</jats:p> |
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| author | Breinig, Sabine, Schiltz, Emile, Fuchs, Georg |
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| description | <jats:title>ABSTRACT</jats:title> <jats:p> Genes involved in the anaerobic metabolism of phenol in the denitrifying bacterium <jats:italic>Thauera aromatica</jats:italic> have been studied. The first two committed steps in this metabolism appear to be phosphorylation of phenol to phenylphosphate by an unknown phosphoryl donor (“phenylphosphate synthase”) and subsequent carboxylation of phenylphosphate to 4-hydroxybenzoate under release of phosphate (“phenylphosphate carboxylase”). Both enzyme activities are strictly phenol induced. Two-dimensional gel electrophoresis allowed identification of several phenol-induced proteins. Based on N-terminal and internal amino acid sequences of such proteins, degenerate oligonucleotides were designed to identify the corresponding genes. A chromosomal DNA segment of about 14 kbp was sequenced which contained 10 genes transcribed in the same direction. These are organized in two adjacent gene clusters and include the genes coding for five identified phenol-induced proteins. Comparison with sequences in the databases revealed the following similarities: the gene products of two open reading frames (ORFs) are each similar to either the central part and N-terminal part of phosphoenolpyruvate synthases. We propose that these ORFs are components of the phenylphosphate synthase system. Three ORFs showed similarity to the <jats:italic>ubiD</jats:italic> gene product, 3-octaprenyl-4-hydroxybenzoate carboxy lyase; UbiD catalyzes the decarboxylation of a 4-hydroxybenzoate analogue in ubiquinone biosynthesis. Another ORF was similar to the <jats:italic>ubiX</jats:italic> gene product, an isoenzyme of UbiD. We propose that (some of) these four proteins are involved in the carboxylation of phenylphosphate. A 700-bp PCR product derived from one of these ORFs cross-hybridized with DNA from different <jats:italic>Thauera</jats:italic> and <jats:italic>Azoarcus</jats:italic> strains, even from those which have not been reported to grow with phenol. One ORF showed similarity to the <jats:italic>mutT</jats:italic> gene product, and three ORFs showed no strong similarities to sequences in the databases. Upstream of the first gene cluster, an ORF which is transcribed in the opposite direction codes for a protein highly similar to the DmpR regulatory protein of <jats:italic>Pseudomonas putida</jats:italic> . DmpR controls transcription of the genes of aerobic phenol metabolism, suggesting a similar regulation of anaerobic phenol metabolism by the putative regulator. </jats:p> |
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| imprint_str_mv | American Society for Microbiology, 2000 |
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| spelling | Breinig, Sabine Schiltz, Emile Fuchs, Georg 0021-9193 1098-5530 American Society for Microbiology Molecular Biology Microbiology http://dx.doi.org/10.1128/jb.182.20.5849-5863.2000 <jats:title>ABSTRACT</jats:title> <jats:p> Genes involved in the anaerobic metabolism of phenol in the denitrifying bacterium <jats:italic>Thauera aromatica</jats:italic> have been studied. The first two committed steps in this metabolism appear to be phosphorylation of phenol to phenylphosphate by an unknown phosphoryl donor (“phenylphosphate synthase”) and subsequent carboxylation of phenylphosphate to 4-hydroxybenzoate under release of phosphate (“phenylphosphate carboxylase”). Both enzyme activities are strictly phenol induced. Two-dimensional gel electrophoresis allowed identification of several phenol-induced proteins. Based on N-terminal and internal amino acid sequences of such proteins, degenerate oligonucleotides were designed to identify the corresponding genes. A chromosomal DNA segment of about 14 kbp was sequenced which contained 10 genes transcribed in the same direction. These are organized in two adjacent gene clusters and include the genes coding for five identified phenol-induced proteins. Comparison with sequences in the databases revealed the following similarities: the gene products of two open reading frames (ORFs) are each similar to either the central part and N-terminal part of phosphoenolpyruvate synthases. We propose that these ORFs are components of the phenylphosphate synthase system. Three ORFs showed similarity to the <jats:italic>ubiD</jats:italic> gene product, 3-octaprenyl-4-hydroxybenzoate carboxy lyase; UbiD catalyzes the decarboxylation of a 4-hydroxybenzoate analogue in ubiquinone biosynthesis. Another ORF was similar to the <jats:italic>ubiX</jats:italic> gene product, an isoenzyme of UbiD. We propose that (some of) these four proteins are involved in the carboxylation of phenylphosphate. A 700-bp PCR product derived from one of these ORFs cross-hybridized with DNA from different <jats:italic>Thauera</jats:italic> and <jats:italic>Azoarcus</jats:italic> strains, even from those which have not been reported to grow with phenol. One ORF showed similarity to the <jats:italic>mutT</jats:italic> gene product, and three ORFs showed no strong similarities to sequences in the databases. Upstream of the first gene cluster, an ORF which is transcribed in the opposite direction codes for a protein highly similar to the DmpR regulatory protein of <jats:italic>Pseudomonas putida</jats:italic> . DmpR controls transcription of the genes of aerobic phenol metabolism, suggesting a similar regulation of anaerobic phenol metabolism by the putative regulator. </jats:p> Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium <i>Thauera aromatica</i> Journal of Bacteriology |
| spellingShingle | Breinig, Sabine, Schiltz, Emile, Fuchs, Georg, Journal of Bacteriology, Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium Thauera aromatica, Molecular Biology, Microbiology |
| title | Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium Thauera aromatica |
| title_full | Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium Thauera aromatica |
| title_fullStr | Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium Thauera aromatica |
| title_full_unstemmed | Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium Thauera aromatica |
| title_short | Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium Thauera aromatica |
| title_sort | genes involved in anaerobic metabolism of phenol in the bacterium <i>thauera aromatica</i> |
| title_unstemmed | Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium Thauera aromatica |
| topic | Molecular Biology, Microbiology |
| url | http://dx.doi.org/10.1128/jb.182.20.5849-5863.2000 |