Literature summary for 1.14.13.24 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.

Search for more literature for 1.14.13.24

Cloned(Commentary)

Cloned (Comment)	Organism
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.

Cloned (Comment)

Organism

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.

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
45000	-	3 * 45000, recombinant His-tagged NarX, SDS-PAGE	Rhodococcus sp.
119000	-	recombinant His-tagged NarX, gel filtration	Rhodococcus sp.

Natural Substrates/ Products (Substrates)

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

Organism

Organism	UniProt	Comment	Textmining
Rhodococcus sp.	E7CYP8	gene narX	-
Rhodococcus sp. NCIMB 12038	E7CYP8	gene narX	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged NarX from Escherichia coli strain Rosetta (DE3) pLysS by nickel affinity chromatography to homogeneity	Rhodococcus sp.

Source Tissue

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

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
3-hydroxybenzoate + NADH + H+ + O2	-	Rhodococcus sp.	2,5-dihydroxybenzoate + NAD+ + H2O	-	?
3-hydroxybenzoate + NADH + H+ + O2	NarX shows activity on 3-hydroxybenzoate rather than on salicylate	Rhodococcus sp.	2,5-dihydroxybenzoate + NAD+ + H2O	-	?
3-hydroxybenzoate + NADH + H+ + O2	-	Rhodococcus sp. NCIMB 12038	2,5-dihydroxybenzoate + NAD+ + H2O	-	?
3-hydroxybenzoate + NADH + H+ + O2	NarX shows activity on 3-hydroxybenzoate rather than on salicylate	Rhodococcus sp. NCIMB 12038	2,5-dihydroxybenzoate + NAD+ + H2O	-	?
3-hydroxybenzoate + NADPH + H+ + O2	-	Rhodococcus sp.	2,5-dihydroxybenzoate + NADP+ + H2O	-	?
3-hydroxybenzoate + NADPH + H+ + O2	-	Rhodococcus sp. NCIMB 12038	2,5-dihydroxybenzoate + NADP+ + H2O	-	?
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.	?	-	?
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	?	-	?

Subunits

Subunits	Comment	Organism
trimer	3 * 45000, recombinant His-tagged NarX, SDS-PAGE	Rhodococcus sp.

Synonyms

Synonyms	Comment	Organism
NarX	-	Rhodococcus sp.

Temperature Optimum [°C]

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

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.4	-	assay at	Rhodococcus sp.

Cofactor

Cofactor	Comment	Organism
FAD	activates by 21%	Rhodococcus sp.
NADH	preferred cofactor	Rhodococcus sp.
NADPH	results in 50% of the activity with NADH	Rhodococcus sp.

General Information

General Information	Comment	Organism
metabolism	NarX is involved in 3-hydroxybenzoate catabolism, organization of the 3HBA-gentisate catabolic cluster, overview	Rhodococcus sp.
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.