Genes in PHT plasmid encoding the initial degradation pathway of phthalate in Pseudomonas putida

Nomura, Yuhta; Nakagawa, Misako; Ogawa, Norio; Harashima, Satoshi; Oshima, Yasuji

doi:10.1016/0922-338x(92)90028-s

Cited by 119 publications

(74 citation statements)

References 29 publications

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“…5). The Pth1 protein was suggested to function either as a positive regulator for the expression of the pth genes or as a phthalate transporter (35). This last assumption was made because of its similarity to the glycerol-3-phosphate transporter (GlpT) of E. coli (15).…”

Section: Resultsmentioning

confidence: 99%

“…Interestingly, hpaA appears to be cotranscribed with the gene hpaX, which encodes a protein of 50,568 Da. A comparative search in the data banks revealed that HpaX is similar (34% identity; 57% similarity) to the protein encoded by the pht1 gene of the phthalate degradative pathway of P. putida (35) (Fig. 5).…”

Section: Resultsmentioning

confidence: 99%

“…Fluorescence-labeled dideoxynucleotide terminators and Taq DNA polymerase were from Applied Biosystems Inc. [␣- 35 S] dCTP and [␣- 32 P]dCTP were from Amersham Corp. Culture media were from Difco. All other chemicals were of the highest grade available and were purchased from Sigma or Merck.…”

Section: Methodsmentioning

confidence: 99%

“…The direct involvement of transport proteins in transcriptional regulation has been well documented (29), and it has been proposed that UhpC controls transcription by bringing inducers into the cell, by directly interacting with the transcriptional regulatory complex, or by a still uncharacterized mechanism (29). On the other hand, it has been shown that some transporters share the same locus with other regulatory proteins, e.g., pth12345 (35), uhpABCT (32), and pcaRKF (21). This could be also the case for hpaX, which appears to form an operon with the hpaA transcriptional regulator.…”

Section: Vol 178 1996mentioning

confidence: 99%

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Molecular characterization of the 4-hydroxyphenylacetate catabolic pathway of Escherichia coli W: engineering a mobile aromatic degradative cluster

1996

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We have determined and analyzed the nucleic acid sequence of a 14,855-bp region that contains the complete gene cluster encoding the 4-hydroxyphenylacetic acid (4-HPA) degradative pathway of Escherichia coli W (ATCC 11105). This catabolic pathway is composed by 11 genes, i.e., 8 enzyme-encoding genes distributed in two putative operons, hpaBC (4-HPA hydroxylase operon) and hpaGEDFHI (meta-cleavage operon); 2 regulatory genes, hpaR and hpaA; and the gene, hpaX, that encodes a protein related to the superfamily of transmembrane facilitators and appears to be cotranscribed with hpaA. Although comparisons with other aromatic catabolic pathways revealed interesting similarities, some of the genes did not present any similarity to their corresponding counterparts in other pathways, suggesting different evolutionary origins. The cluster is flanked by two genes homologous to the cstA (carbon starvation protein) and tsr (serine chemoreceptor) genes of E. coli K-12. A detailed genetic analysis of this region has provided a singular example of how E. coli becomes adapted to novel nutritional sources by the recruitment of a catabolic cassette. Furthermore, the presence of the pac gene in the proximity of the 4-HPA cluster suggests that the penicillin G acylase was a recent acquisition to improve the ability of E. coli W to metabolize a wider range of substrates, enhancing its catabolic versatility. Five repetitive extragenic palindromic sequences that might be involved in transcriptional regulation were found within the cluster. The complete 4-HPA cluster was cloned in plasmid and transposon cloning vectors that were used to engineer E. coli K-12 strains able to grow on 4-HPA. We report here also the in vitro design of new biodegradative capabilities through the construction of a transposable cassette containing the wide substrate range 4-HPA hydroxylase, in order to expand the ortho-cleavage pathway of Pseudomonas putida KT2442 and allow the new recombinant strain to use phenol as the only carbon source.Although most of our current knowledge about the general bacterial metabolic pathways has been derived from the analysis of Escherichia coli, very few data are available about the ability of this microorganism to grow on aromatic compounds other than amino acids. It has been shown that E. coli B, C, and W, but not K-12 strains, are able to degrade 4-hydroxyphenylacetic acid (4-HPA) and homoprotocatechuate (3,4-hydroxyphenylacetate) (HPC) via an inducible, chromosomally encoded meta-cleavage pathway (8, 10).The HPC degradative operon of E. coli C has been partially cloned (25,43), and some of its products have been characterized (16-18, 40-42, 44, 48). In addition, we have previously demonstrated that the first step in the 4-HPA degradation in E. coli W, i.e., the formation of HPC, is catalyzed by a twocomponent aromatic hydroxylase (38,39). This enzyme is encoded by two genes which appear to be part of the same operon (38). The homologous 4-HPA hydroxylase operon of E. coli C has been also cloned and partially sequenced (38...

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Vol 178 1996mentioning

confidence: 99%

See 2 more Smart Citations

Molecular characterization of the 4-hydroxyphenylacetate catabolic pathway of Escherichia coli W: engineering a mobile aromatic degradative cluster

1996

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“…In general, Gram-positive bacteria initially oxidize phthalate to 3,4-dihydro-3,4-dihydroxyphthalate, which is subsequently dehydrogenated and decarboxylated to form protocatechuate [2][3][4]. On the other hand, phthalate degradation in Gramnegative bacteria proceeds through oxygenation and dehydrogenation at carbons 4 and 5 to form 4,5-dihydroxyphthalate, followed by decarboxylation to yield protocatechuate [5].…”

Section: Introductionmentioning

confidence: 99%

Functional Identification of OphR, an IclR Family Transcriptional Regulator Involved in the Regulation of the Phthalate Catabolic Operon in Rhodococcus sp. Strain DK17

et al. 2015

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A putative gene for a transcriptional regulator (ophR) was detected near each copy of the duplicated phthalate-degrading operon of Rhodococcus sp. DK17. Sequence analysis and molecular modeling indicate that OphR belongs to the IclR family of transcriptional regulators and possesses the N-terminal DNA-binding and C-terminal effector-binding domains. DNA-binding assays demonstrate that OphR regulates the phthalate operon by binding to the ophA1-ophR intergenic region.

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Phthalate Dioxygenase Reductase

Ludwig¹,

Ballou²,

Noodleman³

2004

Handbook of Metalloproteins

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Genes in PHT plasmid encoding the initial degradation pathway of phthalate in Pseudomonas putida

Cited by 119 publications

References 29 publications

Molecular characterization of the 4-hydroxyphenylacetate catabolic pathway of Escherichia coli W: engineering a mobile aromatic degradative cluster

Molecular characterization of the 4-hydroxyphenylacetate catabolic pathway of Escherichia coli W: engineering a mobile aromatic degradative cluster

Functional Identification of OphR, an IclR Family Transcriptional Regulator Involved in the Regulation of the Phthalate Catabolic Operon in Rhodococcus sp. Strain DK17

Phthalate Dioxygenase Reductase

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