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Literature summary extracted from

  • Liu, T.T.; Xu, Y.; Liu, H.; Luo, S.; Yin, Y.J.; Liu, S.J.; Zhou, N.Y.
    Functional characterization of a gene cluster involved in gentisate catabolism in Rhodococcus sp. strain NCIMB 12038 (2010), Appl. Microbiol. Biotechnol., 90, 671-678.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.13.11.4 gene narI, DNA and amino acid sequence determination and analysis of the gentisate catabolic gene cluster. Among the 13 complete open reading frames deduced from this fragment, three narIKL encode the enzymes involved in the reactions of gentisate catabolism, of these, NarI is gentisate 1,2-dioxygenase which converts gentisate to maleylpyruvate. Genes narI, narK, and narL genes are transcribed as a single operon only in the presence of naphthalene. Sequence comparisons, overview. Expression in Escherichia coli strain Rosetta (DE3) pLysS Rhodococcus sp.
1.14.13.24 gene narX, DNA and amino acid sequence determination and analysis of the gentisate catabolic gene cluster. Among the 13 complete open reading frames deduced from this fragment, three narIKL encode the enzymes involved in the reactions of gentisate catabolism, of these, NarX is hydroxybenzoate 6-monooxygenase which converts 3-hydroxybenzoate to 2,5-dihydroxybenzoate. The narX gene is divergently transcribed with narIKL. Sequence comparisons, overview. Expression in of the His-tagged NarX in Escherichia coli strain Rosetta (DE3) pLysS Rhodococcus sp.
5.2.1.4 expressed in Escherichia coli Rosetta (DE3) pLysS cells Rhodococcus sp.

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.14.13.24 45000
-
 3 * 45000, recombinant His-tagged NarX, SDS-PAGE Rhodococcus sp.
1.14.13.24 119000
-
 recombinant His-tagged NarX, gel filtration Rhodococcus sp.
5.2.1.4 29000
-
 gel filtration Rhodococcus sp.
5.2.1.4 30000
-
 1 * 30000, SDS-PAGE Rhodococcus sp.

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.13.11.4 2,5-dihydroxybenzoate + O2 Rhodococcus sp.
-
 maleylpyruvate
-
 ?
1.13.11.4 2,5-dihydroxybenzoate + O2 Rhodococcus sp. NCIMB 12038
-
 maleylpyruvate
-
 ?
1.14.13.24 3-hydroxybenzoate + NADH + H+ + O2 Rhodococcus sp. NarX shows activity on 3-hydroxybenzoate rather than on salicylate 2,5-dihydroxybenzoate + NAD+ + H2O
-
 ?
1.14.13.24 3-hydroxybenzoate + NADH + H+ + O2 Rhodococcus sp. NCIMB 12038 NarX shows activity on 3-hydroxybenzoate rather than on salicylate 2,5-dihydroxybenzoate + NAD+ + H2O
-
 ?
1.14.13.24 3-hydroxybenzoate + NADPH + H+ + O2 Rhodococcus sp.
-
 2,5-dihydroxybenzoate + NADP+ + H2O
-
 ?
1.14.13.24 3-hydroxybenzoate + NADPH + H+ + O2 Rhodococcus sp. NCIMB 12038
-
 2,5-dihydroxybenzoate + NADP+ + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.13.11.4 Rhodococcus sp. E7CYQ0 gene narI
-
1.13.11.4 Rhodococcus sp. NCIMB 12038 E7CYQ0 gene narI
-
1.14.13.24 Rhodococcus sp. E7CYP8 gene narX
-
1.14.13.24 Rhodococcus sp. NCIMB 12038 E7CYP8 gene narX
-
5.2.1.4 Rhodococcus sp.
-
-
-
5.2.1.4 Rhodococcus sp. NCIMB 12038
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.13.24 recombinant His-tagged NarX from Escherichia coli strain Rosetta (DE3) pLysS by nickel affinity chromatography to homogeneity Rhodococcus sp.
5.2.1.4 Ni-NTA agarose column chromatography Rhodococcus sp.

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.13.11.4 additional information strain NCIMB 12038 utilizes naphthalene as a sole source of carbon and energy Rhodococcus sp.
-
1.14.13.24 additional information strain NCIMB 12038 utilizes naphthalene as a sole source of carbon and energy Rhodococcus sp.
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.13.11.4 145
-
 recombinant enzyme, pH 7.4, 30°C Rhodococcus sp.
1.14.13.24 5.8
-
 purified recombinant His-tagged NarX, pH 7.4, 30°C Rhodococcus sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.13.11.4 2,5-dihydroxybenzoate + O2
-
 Rhodococcus sp. maleylpyruvate
-
 ?
1.13.11.4 2,5-dihydroxybenzoate + O2
-
 Rhodococcus sp. NCIMB 12038 maleylpyruvate
-
 ?
1.14.13.24 3-hydroxybenzoate + NADH + H+ + O2
-
 Rhodococcus sp. 2,5-dihydroxybenzoate + NAD+ + H2O
-
 ?
1.14.13.24 3-hydroxybenzoate + NADH + H+ + O2 NarX shows activity on 3-hydroxybenzoate rather than on salicylate Rhodococcus sp. 2,5-dihydroxybenzoate + NAD+ + H2O
-
 ?
1.14.13.24 3-hydroxybenzoate + NADH + H+ + O2
-
 Rhodococcus sp. NCIMB 12038 2,5-dihydroxybenzoate + NAD+ + H2O
-
 ?
1.14.13.24 3-hydroxybenzoate + NADH + H+ + O2 NarX shows activity on 3-hydroxybenzoate rather than on salicylate Rhodococcus sp. NCIMB 12038 2,5-dihydroxybenzoate + NAD+ + H2O
-
 ?
1.14.13.24 3-hydroxybenzoate + NADPH + H+ + O2
-
 Rhodococcus sp. 2,5-dihydroxybenzoate + NADP+ + H2O
-
 ?
1.14.13.24 3-hydroxybenzoate + NADPH + H+ + O2
-
 Rhodococcus sp. NCIMB 12038 2,5-dihydroxybenzoate + NADP+ + H2O
-
 ?
1.14.13.24 additional information the purified H6-NarX protein shows no detectable activity against the 3HBA structural analogues, including 2-hydroxybenzoate, i.e. salicylate, 4-hydroxybenzoate, or 4-substituted derivatives (4-CH3, 4-NH2, and 4-NO2) of 3-hydroxybenzoate Rhodococcus sp. ?
-
 ?
1.14.13.24 additional information the purified H6-NarX protein shows no detectable activity against the 3HBA structural analogues, including 2-hydroxybenzoate, i.e. salicylate, 4-hydroxybenzoate, or 4-substituted derivatives (4-CH3, 4-NH2, and 4-NO2) of 3-hydroxybenzoate Rhodococcus sp. NCIMB 12038 ?
-
 ?
5.2.1.4 Maleylpyruvate
-
 Rhodococcus sp. Fumarylpyruvate
-
 ?
5.2.1.4 Maleylpyruvate
-
 Rhodococcus sp. NCIMB 12038 Fumarylpyruvate
-
 ?

Subunits

EC Number Subunits Comment Organism
1.14.13.24 trimer 3 * 45000, recombinant His-tagged NarX, SDS-PAGE Rhodococcus sp.
5.2.1.4 monomer 1 * 30000, SDS-PAGE Rhodococcus sp.

Synonyms

EC Number Synonyms Comment Organism
1.13.11.4 GDO
-
 Rhodococcus sp.
1.13.11.4 NarI
-
 Rhodococcus sp.
1.14.13.24 NarX
-
 Rhodococcus sp.
5.2.1.4 mycothiol-dependent maleylpyruvate isomerase
-
 Rhodococcus sp.
5.2.1.4 NarL
-
 Rhodococcus sp.

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.13.11.4 30
-
 assay at Rhodococcus sp.
1.14.13.24 30
-
 assay at Rhodococcus sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.13.11.4 7.4
-
 assay at Rhodococcus sp.
1.14.13.24 7.4
-
 assay at Rhodococcus sp.

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.13.24 FAD activates by 21% Rhodococcus sp.
1.14.13.24 NADH preferred cofactor Rhodococcus sp.
1.14.13.24 NADPH results in 50% of the activity with NADH Rhodococcus sp.
5.2.1.4 mycothiol dependent on Rhodococcus sp.

General Information

EC Number General Information Comment Organism
1.13.11.4 metabolism narI is involved in salicylate, i.e. 3-hydroxybenzoate, and gentisate catabolism, organization of the 3HBA-gentisate catabolic cluster, overview Rhodococcus sp.
1.13.11.4 additional information strain NCIMB 12038 utilizes naphthalene as a sole source of carbon and energy, and degrades naphthalene via salicylate and gentisate. The strain can also use salicylate, i.e. 3-hydroxybenzoate, as sole source of energy and carbon Rhodococcus sp.
1.14.13.24 metabolism NarX is involved in 3-hydroxybenzoate catabolism, organization of the 3HBA-gentisate catabolic cluster, overview Rhodococcus sp.
1.14.13.24 additional information strain NCIMB 12038 utilizes naphthalene as a sole source of carbon and energy, and degrades naphthalene via salicylate and gentisate. The strain can also use salicylate, i.e. 3-hydroxybenzoate, as sole source of energy and carbon Rhodococcus sp.