Information on EC 1.14.13.166 - 4-nitrocatechol 4-monooxygenase

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The enzyme appears in viruses and cellular organisms

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EC NUMBER
COMMENTARY hide
1.14.13.166
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RECOMMENDED NAME
GeneOntology No.
4-nitrocatechol 4-monooxygenase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
4-nitrocatechol + NAD(P)H + H+ + O2 = 2-hydroxy-1,4-benzoquinone + nitrite + NAD(P)+ + H2O
show the reaction diagram
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
4-nitrophenol degradation II
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SYSTEMATIC NAME
IUBMB Comments
4-nitrocatechol,NAD(P)H:oxygen 4-oxidoreductase (4-hydroxylating, nitrite-forming)
Contains FAD. The enzyme catalyses the oxidation of 4-nitrocatechol with the concomitant removal of the nitro group as nitrite. Forms a two-component system with a flavoprotein reductase [1]. The enzymes from the bacteria Lysinibacillus sphaericus JS905 and Rhodococcus sp. strain PN1 were shown to also catalyse EC 1.14.13.29, 4-nitrophenol 2-monooxygenase [1,2] while the enzyme from Pseudomonas sp. WBC-3 was shown to also catalyse EC 1.14.13.167, 4-nitrophenol 4-monooxygenase [3].
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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A7YVV2
UniProt
Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
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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
2,4,5-trichlorophenol + NADH + H+ + O2
2,5-dichlorohydroquinone + 5-chloro-2-hydroxyquinol + ? + NAD+ + H2O
show the reaction diagram
A7YVV2
poor substrate
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?
2,4,6-trichlorophenol + NADH + H+ + O2
2,6-dichlorohydroquinone + 6-chloro-2-hydroxyquinol + ? + NAD+ + H2O
show the reaction diagram
A7YVV2
poor substrate
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?
2,4-dinitrophenol + NADH + H+ + O2
2-nitrohydroquinone + nitrite + NAD+ + H2O
show the reaction diagram
A7YVV2
poor substrate
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?
3-chlorophenol + NADH + H+ + O2
? + NAD+ + H2O
show the reaction diagram
A7YVV2
poor substrate
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?
3-cresol + NADH + H+ + O2
? + NAD+ + H2O
show the reaction diagram
A7YVV2
poor substrate
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?
3-nitrophenol + NADH + H+ + O2
3-nitrohydroquinone + NAD+ + H2O
show the reaction diagram
A7YVV2
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?
4-chlorocatechol + NADH + H+ + O2
1,2,4-trihydroxybenzene + ? + NAD+ + H2O
show the reaction diagram
A7YVV2
good substrate
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?
4-chlorophenol + NADH + H+ + O2
1,2,4-trihydroxybenzene + 2-hydroxy-1,4-benzoquinone + ? + NAD+ + H2O
show the reaction diagram
A7YVV2
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?
4-chlororesorcinol + NADH + H+ + O2
1,2,4-trihydroxybenzene + ? + NAD+ + H2O
show the reaction diagram
A7YVV2
good substrate
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?
4-cresol + NADH + H+ + O2
4-hydroxy-4-methyl-2,5-cyclohexadien-1-one + NAD+ + H2O
show the reaction diagram
A7YVV2
poor substrate
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?
4-methylcatechol + NADH + H+ + O2
? + NAD+ + H2O
show the reaction diagram
A7YVV2
poor substrate
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?
4-nitrocatechol + NAD(P)H + H+ + O2
1,2,4-trihydroxybenzene + nitrite + NAD(P)+ + H2O
show the reaction diagram
4-nitrocatechol + NAD(P)H + H+ + O2
2-hydroxy-1,4-benzoquinone + nitrite + NAD(P)+ + H2O
show the reaction diagram
4-nitrocatechol + NADH + H+ + O2
1,2,4-trihydroxybenzene + nitrite + NAD+ + H2O
show the reaction diagram
4-nitrocatechol + NADH + H+ + O2
2-hydroxy-1,4-benzoquinone + nitrite + NAD+ + H2O
show the reaction diagram
4-nitrocatechol + NADPH + H+ + O2
1,2,4-trihydroxybenzene + nitrite + NADP+ + H2O
show the reaction diagram
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monooxygenation, the enzyme completely degrades 0.1 mM 4-nitrocatechol in 80 min
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?
4-nitrocatechol + NADPH + H+ + O2
?
show the reaction diagram
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?
4-nitrophenol + NADH + H+ + O2
2-hydroxy-1,4-benzoquinone + 1,2,4-trihydroxybenzene + nitrite + NAD+ + H2O
show the reaction diagram
A7YVV2
best substrate
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?
phenol + NADH + H+ + O2
hydroquinone + ? + NAD+ + H2O
show the reaction diagram
A7YVV2
poor substrate
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?
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mg2+
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magnesium ions enhance the enzyme activity
additional information
addition of Mg2+ or Mn2+ ions has no effect on the activity
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
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Methimazole
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58% inhibition at 0.5 mM
additional information
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alpha-naphthoflavone, miconazole, and metyrapone at 0.5 mM have very little effect (less than 5% inhibition) on enzyme activity
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0013
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cell extract, using 4-nitrocatechol as substrate, at pH 8.0 and 30°C; cell extract, using 4-nitrophenol as substrate, at pH 8.0 and 30°C
0.038
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after 28fold purification, using 4-nitrocatechol as substrate, at pH 8.0 and 30°C
0.04
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after 32fold purification, using 4-nitrophenol as substrate, at pH 8.0 and 30°C
1.9
at pH 7.0 and 30°C
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8
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in 50 mM Tris-HCl
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
46600
x * 46600, calculated from amino acid sequence
47000
1 * 47000, His-tagged enzyme, gel filtration
59933
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x * 59933, calculated from amino acid sequence
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SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
1 * 47000, His-tagged enzyme, gel filtration
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Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
sitting drop vapor diffusion method, using 20 mM HEPES pH 7.0, 15% (w/v) PEG 4000 and 15% (v/v) 2-propanol
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, without glycerol, rapid inactivation
-70°C, in 50% (v/v) glycerol, 7 days, 70% loss of activity
0°C, extracts prepared without added FAD, 48 h, 52% loss of activity
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0°C, storage medium, no loss of activity after several hours, 30% loss of activity after 12 h, 80% loss of activity after 40 h
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DEAE-Sepharose column chromatography, Q-Sepharose column chromatography, and Sephacryl S-300 gel filtration
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Ni-NTA agarose column chromatography
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Ni-NTA agarose column chromatography and Sephadex G-100 gel filtration
Ni-NTA column chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli BL21(DE3)pLysS cells
A7YVV2
His-tagged enzyme is expressed in Escherichia coli BL21(DE3) cells
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
enzyme expression can be induced by 4-nitrophenol but not 4-nitrocatechol
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LINKS TO OTHER DATABASES (specific for EC-Number 1.14.13.166)
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)