γ-Resorcylate Catabolic-Pathway Genes in the Soil Actinomycete Rhodococcus jostii RHA1

Kasai, Daisuke; Araki, Naoto; Motoi, Kota; Yoshikawa, Shota; Iino, Toju; Imai, Shunsuke; Masai, Eiji; Fukuda, Masao

doi:10.1128/aem.02422-15

Cited by 12 publications

(10 citation statements)

References 49 publications

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“…MTP-10005 (11), Rhizobium radiobacter WU-0108 (12), Agrobacterium tumefaciens IAM12048 (13), Pandoraea sp. 12B-2 (14) and Rhodococcus jostii RHA1 (15). In the γ-resorcylate catabolic pathway identified in Rhizobium sp.…”

Section: Introductionmentioning

confidence: 99%

Mechanism and Structure of γ-Resorcylate Decarboxylase

et al. 2017

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γ-Resorcylate decarboxylase (γ-RSD) has evolved to catalyze the reversible decarboxylation of 2,6-dihydroxybenzoate to resorcinol in a nonoxidative fashion. This enzyme is of significant interest because of its potential for the production of γ-resorcylate and other benzoic acid derivatives under environmentally sustainable conditions. Kinetic constants for the decarboxylation of 2,6-dihydroxybenzoate catalyzed by γ-RSD from Polaromonas sp. JS666 are reported, and the enzyme is shown to be active with 2,3-dihydroxybenzoate, 2,4,6-trihydroxybenzoate, and 2,6-dihydroxy-4-methylbenzoate. The three-dimensional structure of γ-RSD with the inhibitor 2-nitroresorcinol (2-NR) bound in the active site is reported. 2-NR is directly ligated to a Mn bound in the active site, and the nitro substituent of the inhibitor is tilted significantly from the plane of the phenyl ring. The inhibitor exhibits a binding mode different from that of the substrate bound in the previously determined structure of γ-RSD from Rhizobium sp. MTP-10005. On the basis of the crystal structure of the enzyme from Polaromonas sp. JS666, complementary density functional calculations were performed to investigate the reaction mechanism. In the proposed reaction mechanism, γ-RSD binds 2,6-dihydroxybenzoate by direct coordination of the active site manganese ion to the carboxylate anion of the substrate and one of the adjacent phenolic oxygens. The enzyme subsequently catalyzes the transfer of a proton to C1 of γ-resorcylate prior to the actual decarboxylation step. The reaction mechanism proposed previously, based on the structure of γ-RSD from Rhizobium sp. MTP-10005, is shown to be associated with high energies and thus less likely to be correct.

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Section: Introductionmentioning

confidence: 99%

Mechanism and Structure of γ-Resorcylate Decarboxylase

et al. 2017

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“…PN1 [8], as an aromatic catabolic pathway, and a hydroxyquinol degradation gene cluster has been observed in Cupriavidus necator JMP134 [14]. This pathway has been implicated in degradation of 4-nitrophenol in Arthrobacter chlorophenolicus A6 [15], and in -resorcylate catabolism in R. jostii RHA1 [9]. Following our initial observation that a pcaHG gene deletion mutant of R. jostii RHA1 was able to grow on minimal media containing protocatechuic acid, we hypothesised that protocatechuic acid could be converted via flavin-dependent mono-oxygenase (ro01860) and a decarboxylase (ro01859) into hydroxyquinol.…”

Section: Discussionmentioning

confidence: 99%

“…Only a small amount of hydroxylation of catechol by mono-oxygenase Ro01860 alone was observed, implying that the two enzymes are more active in combination, perhaps forming a complex. Kasai et al have shown that the R. jostii ro01859 and ro01860 gene products are able to convert 2,6-dihydroxybenzoic acid to hydroxyquinol: Ro01859 decarboxylates -resorcylate to resorcinol, and Ro01860 converts resorcinol to hydroxyquinol [9].…”

Section: Discussionmentioning

confidence: 99%

“…PN1 [8]. Kasai et al have implicated R. jostii RHA1 genes ro01857-ro01861 in -resorcylate catabolism, via decarboxylation of gammaresorcylate (2,6-dihydroxybenzoic acid) and hydroxylation of resorcinol to form hydroxyquinol [9]. Transcription of genes ro01857-ro01860 was shown to be regulated by gene regulator TsdR, to which gamma-resorcylate binds as an effector [9].…”

Section: Introductionmentioning

confidence: 99%

“…Kasai et al have implicated R. jostii RHA1 genes ro01857-ro01861 in -resorcylate catabolism, via decarboxylation of gammaresorcylate (2,6-dihydroxybenzoic acid) and hydroxylation of resorcinol to form hydroxyquinol [9]. Transcription of genes ro01857-ro01860 was shown to be regulated by gene regulator TsdR, to which gamma-resorcylate binds as an effector [9]. The oxidative decarboxylation of vanillic acid to 2methoxyhydroquinone has been reported in extracts of Sporotrichum pulverulentum [10], hence the oxidative decarboxylation of protocatechuic acid to form hydroxyquinol has some biochemical precedent.…”

Section: Introductionmentioning

confidence: 99%

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The Hydroxyquinol Degradation Pathway in Rhodococcus jostii RHA1 and Agrobacterium Species Is an Alternative Pathway for Degradation of Protocatechuic Acid and Lignin Fragments

Spence

Scott

Dumond

et al. 2020

Appl Environ Microbiol

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Deletion of the pcaHG genes encoding protocatechuate 3,4-dioxygenase in Rhodococcus jostii RHA1 gives a gene deletion strain still able to grow on protocatechuic acid as sole carbon source, indicating a second degradation pathway for protocatechuic acid. Metabolite analysis of wild-type R. jostii RHA1 grown on media containing vanillin or protocatechuic acid indicated the formation of hydroxyquinol (benzene-1,2,4-triol) as a downstream product. Gene cluster ro01857-ro01860 in Rhodococcus jostii RHA1 contains genes encoding hydroxyquinol 1,2-dioxygenase and maleylacetate reductase for degradation of hydroxyquinol but also putative mono-oxygenase (ro01860) and putative decarboxylase (ro01859) genes, and a similar gene cluster is found in the genome of lignin-degrading Agrobacterium sp.. Recombinant R. jostii mono-oxygenase and decarboxylase enzymes were in combination found to convert protocatechuic acid to hydroxyquinol. Hence an alternative pathway for degradation of protocatechuic acid via oxidative decarboxylation to hydroxyquinol is proposed. Importance There is a well-established paradigm for degradation of protocatechuic acid via the β-ketoadipate pathway in a range of soil bacteria. In this study we have found the existence of a second pathway for degradation of protocatechuic acid in Rhodococcus jostii RHA1, via hydroxyquinol (benzene-1,2,4-triol), which establishes a metabolic link between protocatechuic acid and hydroxyquinol. The presence of this pathway in a lignin-degrading Agrobacterium sp. strain suggests a possible involvement of the hydroxyquinol pathway in metabolism of lignin degradation fragments.

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