Information on EC 1.14.13.172 - salicylate 5-hydroxylase

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The enzyme appears in viruses and cellular organisms

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EC NUMBER
COMMENTARY hide
1.14.13.172
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RECOMMENDED NAME
GeneOntology No.
salicylate 5-hydroxylase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
salicylate + NADH + H+ + O2 = 2,5-dihydroxybenzoate + NAD+ + H2O
show the reaction diagram
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Naphthalene degradation
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Microbial metabolism in diverse environments
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salicylate glucosides biosynthesis II
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salicylate degradation II
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SYSTEMATIC NAME
IUBMB Comments
salicylate,NADH:oxygen oxidoreductase (5-hydroxylating)
This enzyme, which was characterized from the bacterium Ralstonia sp. U2, comprises a multicomponent system, containing a reductase that is an iron-sulfur flavoprotein (FAD; EC 1.18.1.7, ferredoxin---NAD(P)+ reductase), an iron-sulfur oxygenase, and ferredoxin.
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
large oxygenase component and small oxagenase component
O52379 and O52380
UniProt
Manually annotated by BRENDA team
large oxygenase component and small oxagenase component
O52379 and O52380
UniProt
Manually annotated by BRENDA team
large oxygenase component and small oxagenase component
O52379 and O52380
UniProt
Manually annotated by BRENDA team
large oxygenase component and small oxagenase component
O52379 and O52380
UniProt
Manually annotated by BRENDA team
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2,4-dihydroxybenzoate + NADH + H+ + O2
?
show the reaction diagram
2,6-dihydroxybenzoate + NADH + H+ + O2
?
show the reaction diagram
2-nitrophenol + NADH + H+ + O2
?
show the reaction diagram
salicylate + NADH + H+ + O2
2,5-dihydroxybenzoate + NAD+ + H2O
show the reaction diagram
salicylate + NADPH + H+ + O2
2,5-dihydroxybenzoate + NADP+ + H2O
show the reaction diagram
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
salicylate + NADH + H+ + O2
2,5-dihydroxybenzoate + NAD+ + H2O
show the reaction diagram
salicylate + NADPH + H+ + O2
2,5-dihydroxybenzoate + NADP+ + H2O
show the reaction diagram
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADPH
[2Fe-2S]-center
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Iron
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the enzyme contains one Rieske-type [2Fe-2S] cluster and 7.5 g iron per mol enzyme
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.9398
2,6-dihydroxybenzoate
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in 50 mM MES buffer (pH 6.4), at 22°C
0.3284
2-Nitrophenol
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in 50 mM MES buffer (pH 6.4), at 22°C
0.05976
NADH
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in 50 mM MES buffer (pH 6.4), at 22°C
0.05641
NADPH
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in 50 mM MES buffer (pH 6.4), at 22°C
0.1071
salicylate
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in 50 mM MES buffer (pH 6.4), at 22°C
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.051
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in 50 mM MES buffer, pH 6.4, at 30°C
107.1
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after 124fold purification, in 50 mM MES buffer (pH 6.4), at 22°C
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20000
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3 * 52000 + 3 * 20000, His6-tagged enzyme, SDS-PAGE
45000
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4 * 45000, SDS-PAGE
48799
O52379 and O52380
x * 48799 + x * 18800, calculated from amino acid sequence
51000
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3 * 51000 + 3 * 18800, His6-tagged enzyme, calculated from amino acid sequence
52000
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3 * 52000 + 3 * 20000, His6-tagged enzyme, SDS-PAGE
180000
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gel filtration
195200
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gel filtration
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SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterohexamer
homotetramer
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
the three-component Fe-S protein complex (NagAaGHAb) of the enzyme is purified by HisTrap column chromatography and Sephadex G-25 gel filtration
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
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expressed in Escherichia coli BL21(DE3)/pLysS cells
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expressed in Escherichia coli DH5alpha cells
O52379 and O52380
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LINKS TO OTHER DATABASES (specific for EC-Number 1.14.13.172)
ExplorEnz
ExPASy
KEGG
MetaCyc
SABIO-RK
NCBI: PubMed, Protein, Nucleotide, Structure, Genome, OMIM
IUBMB Enzyme Nomenclature
PROSITE Database of protein families and domains
InterPro (database of protein families, domains and functional sites)