Gentisate pathway in Salmonella typhimurium: metabolism of m-hydroxybenzoate and gentisate

Goetz, Frederick E.; Harmuth, Lorna J.

doi:10.1016/0378-1097(92)90361-q

Cited by 12 publications

(12 citation statements)

References 23 publications

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“…Salicylate degradation has been studied in Pseudomonas spp., and catechol and gentisate are key intermediates in the two known pathways (9,24,27,65). Salicylate is converted to catechol and gentisate by salicylate hydroxylase (4,6) and salicylate 5-hydroxylase (22,27), respectively.…”

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confidence: 99%

Novel Pathway of Salicylate Degradation by Streptomyces sp. Strain WA46

Ishiyama

Vujaklija

Davies

2004

Appl Environ Microbiol

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A novel salicylate-degrading Streptomyces sp., strain WA46, was identified by UV fluorescence on solid minimal medium containing salicylate; trace amounts of gentisate were detected by high-pressure liquid chromatography when strain WA46 was grown with salicylate. PCR amplification of WA46 DNA with degenerate primers for gentisate 1,2-dioxygenase (GDO) genes produced an amplicon of the expected size. Sequential PCR with nested GDO primers was then used to identify a salicylate degradation gene cluster in a plasmid library of WA46 chromosomal DNA. The nucleotide sequence of a 13.5-kb insert in recombinant plasmid pWD1 (which was sufficient for the complete degradation of salicylate) showed that nine putative open reading frames (ORFs) (sdgABCDEFGHR) were involved. Plasmid pWD1 derivatives disrupted in each putative gene were transformed into Streptomyces lividans TK64. Disruption of either sdgA or sdgC blocked salicylate degradation; constructs lacking sdgD accumulated gentisate. Cell extracts from Escherichia coli DH5␣ transformants harboring pUC19 that expressed each of the sdg ORFs showed that conversions of salicylate to salicylylcoenzyme A (CoA) and salicylyl-CoA to gentisyl-CoA required SdgA and SdgC, respectively. SdgA required CoA and ATP as cofactors, while NADH was required for SdgC activity; SdgC was identified as salicylyl-CoA 5-hydroxylase. Gentisyl-CoA underwent spontaneous cleavage to gentisate and CoA. SdgA behaved as a salicylyl-CoA ligase despite showing amino acid sequence similarity to an AMP-ligase. SdgD was identified as a GDO. These results suggest that Streptomyces sp. strain WA46 degrades salicylate by a novel pathway via a CoA derivative. Two-dimensional polyacrylamide gel electrophoresis and reverse transcriptase-PCR studies indicated that salicylate induced expression of the sdg cluster.

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confidence: 99%

Novel Pathway of Salicylate Degradation by Streptomyces sp. Strain WA46

Ishiyama

Vujaklija

Davies

2004

Appl Environ Microbiol

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“…However, gentisate and substituted gentisates serve as key intermediates in the aerobic pathways for the metabolism of a large number of aromatic compounds, including 3-hydroxybenzoate (15,24), substituted phenols (8,23,39), salicylate (37,40), 3,6-dichloro-2-methoxybenzoate (53), and naphthalene (14,17,34). Ring cleavage of gentisate is catalyzed by gentisate 1,2-dioxygenase (GDO; EC1.13.11.4) to form maleylpyruvate (26).…”

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nag Genes of Ralstonia (Formerly Pseudomonas ) sp. Strain U2 Encoding Enzymes for Gentisate Catabolism

2001

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Ralstonia sp. strain U2 metabolizes naphthalene via gentisate to central metabolites. We have cloned and sequenced a 21.6-kb region spanning the nag genes. Upstream of the pathway genes are nagY, homologous to chemotaxis proteins, and nagR, a regulatory gene of the LysR family. Divergently transcribed from nagR are the genes for conversion of naphthalene to gentisate (nagAaGHAbAcAdBFCQED) (S. L. Fuenmayor, M. Wild, A. L. Boyes, and P. A. Williams, J. Bacteriol. 180:2522-2530, 1998), which except for the insertion of nagGH, encoding the salicylate 5-hydroxylase, are homologous to and in the same order as the genes in the classical upper pathway operon described for conversion of naphthalene to salicylate found in the NAH7 plasmid of Pseudomonas putida PpG7. Downstream of nahD is a cluster of genes (nagJIKLMN) which are probably cotranscribed with nagAaGHAbAcAdBFCQED as a single large operon. By cloning into expression vectors and by biochemical assays, three of these genes (nagIKL) have been shown to encode the enzymes involved in the further catabolism of gentisate to fumarate and pyruvate. NagI is a gentisate 1,2-dioxygenase which converts gentisate to maleylpyruvate and is also able to catalyze the oxidation of some substituted gentisates. NagL is a reduced glutathione-dependent maleylpyruvate isomerase catalyzing the isomerization of maleylpyruvate to fumarylpyruvate. NagK is a fumarylpyruvate hydrolase which hydrolyzes fumarylpyruvate to fumarate and pyruvate. The three other genes (nagJMN) have also been cloned and overexpressed, but no biochemical activities have been attributed to them. NagJ is homologous to a glutathione S-transferase, and NagM and NagN are proteins homologous to each other and to other proteins of unknown function. Downstream of the operon is a partial sequence with homology to a transposase.

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“…2,5-Dihydroxybenzoate is an intermediate in the gentisate pathway of aromatic degradation (Fig. 1) ; although this pathway has been identified in a variety of soil bacteria (Dagley, 1975) and in the enteric bacteria Klebsiella pnenmoniae (Jones & Cooper, 1990) and Salmonella t_yphimtrrinm (Goetz & Harmuth, 1992) little is known about the pathway genes.…”

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confidence: 99%

In vitro formation of a catabolic plasmid carrying Klebsiella pneumoniae DNA that allows growth of Escherichia coli K-12 on 3-hydroxybenzoate

Robson

Parrott²,

Cooper³

1996

Microbiology

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The four enzymes needed to convert 3-hydroxybenzoate to pyruvate and fumarate via the gentisate pathway, as well as a putative positive regulator protein, were encoded on an 8 kb Sphl fragment of Klebsiella pneumoniae DNA. The five genes were clustered in the order regulator-gentisate dioxygenase-furnarylpyruvate hydrolase-3-hydroxybenzoate monooxygenase-maleylpyruvate isomerase (mhbRDHMI ), with the catabolic genes transcribed in the dioxygenase to isomerase direction. 2-Hydroxybenzoate was found to be a non-metabolizable inducer analogue for the mhb genes, supporting the view that gentisate rather than maleylpyruvate was the physiological inducer. The plasmid pNDR2O encoding the full gentisate catabolic pathway endowed €scherichia coli with the ability to grow on 3-hydroxybenzoate but the host cell appeared to be responsible for substrate uptake.

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Gentisate pathway in Salmonella typhimurium: metabolism of m-hydroxybenzoate and gentisate

Cited by 12 publications

References 23 publications

Novel Pathway of Salicylate Degradation by Streptomyces sp. Strain WA46

Novel Pathway of Salicylate Degradation by Streptomyces sp. Strain WA46

nag Genes of Ralstonia (Formerly Pseudomonas ) sp. Strain U2 Encoding Enzymes for Gentisate Catabolism

In vitro formation of a catabolic plasmid carrying Klebsiella pneumoniae DNA that allows growth of Escherichia coli K-12 on 3-hydroxybenzoate

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