Information on EC 1.14.13.166 - 4-nitrocatechol 4-monooxygenase

for references in articles please use BRENDA:EC1.14.13.166
Word Map on EC 1.14.13.166
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:

The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.13.166
-
RECOMMENDED NAME
GeneOntology No.
4-nitrocatechol 4-monooxygenase
-
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
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
4-nitrophenol degradation II
-
-
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].
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
no activity in Rhodococcus imtechensis
strain RKJ300, DSM 45091
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
physiological function
additional information
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
poor substrate
-
-
?
2,4,6-trichlorophenol + NADH + H+ + O2
2,6-dichlorohydroquinone + 6-chloro-2-hydroxyquinol + ? + NAD+ + H2O
show the reaction diagram
poor substrate
-
-
?
2,4-dinitrophenol + NADH + H+ + O2
2-nitrohydroquinone + nitrite + NAD+ + H2O
show the reaction diagram
poor substrate
-
-
?
3-chlorophenol + NADH + H+ + O2
? + NAD+ + H2O
show the reaction diagram
poor substrate
-
-
?
3-cresol + NADH + H+ + O2
? + NAD+ + H2O
show the reaction diagram
poor substrate
-
-
?
3-nitrophenol + NADH + H+ + O2
3-nitrohydroquinone + NAD+ + H2O
show the reaction diagram
-
-
-
?
4-chlorocatechol + NADH + H+ + O2
1,2,4-trihydroxybenzene + ? + NAD+ + H2O
show the reaction diagram
good substrate
-
-
?
4-chlorophenol + NADH + H+ + O2
1,2,4-trihydroxybenzene + 2-hydroxy-1,4-benzoquinone + ? + NAD+ + H2O
show the reaction diagram
-
-
-
?
4-chlororesorcinol + NADH + H+ + O2
1,2,4-trihydroxybenzene + ? + NAD+ + H2O
show the reaction diagram
good substrate
-
-
?
4-cresol + NADH + H+ + O2
4-hydroxy-4-methyl-2,5-cyclohexadien-1-one + NAD+ + H2O
show the reaction diagram
poor substrate
-
-
?
4-methylcatechol + NADH + H+ + O2
? + NAD+ + H2O
show the reaction diagram
poor substrate
-
-
?
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
-
monooxygenation, the enzyme completely degrades 0.1 mM 4-nitrocatechol in 80 min
-
-
?
4-nitrocatechol + NADPH + H+ + O2
?
show the reaction diagram
-
-
-
?
4-nitrophenol + NADH + H+ + O2
2-hydroxy-1,4-benzoquinone + 1,2,4-trihydroxybenzene + nitrite + NAD+ + H2O
show the reaction diagram
best substrate
-
-
?
phenol + NADH + H+ + O2
hydroquinone + ? + NAD+ + H2O
show the reaction diagram
poor substrate
-
-
?
additional information
?
-
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
4-nitrocatechol + NAD(P)H + H+ + O2
2-hydroxy-1,4-benzoquinone + nitrite + NAD(P)+ + H2O
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mg2+
-
magnesium ions enhance the enzyme activity
additional information
addition of Mg2+ or Mn2+ ions has no effect on the activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
-
-
Methimazole
-
58% inhibition at 0.5 mM
additional information
-
alpha-naphthoflavone, miconazole, and metyrapone at 0.5 mM have very little effect (less than 5% inhibition) on enzyme activity
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0013
-
cell extract, using 4-nitrocatechol as substrate, at pH 8.0 and 30C; cell extract, using 4-nitrophenol as substrate, at pH 8.0 and 30C
0.038
-
after 28fold purification, using 4-nitrocatechol as substrate, at pH 8.0 and 30C
0.04
-
after 32fold purification, using 4-nitrophenol as substrate, at pH 8.0 and 30C
1.9
at pH 7.0 and 30C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8
-
in 50 mM Tris-HCl
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
46600
x * 46600, calculated from amino acid sequence
47000
1 * 47000, His-tagged enzyme, gel filtration
59933
-
x * 59933, calculated from amino acid sequence
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
1 * 47000, His-tagged enzyme, gel filtration
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
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, without glycerol, rapid inactivation
-70C, in 50% (v/v) glycerol, 7 days, 70% loss of activity
0C, extracts prepared without added FAD, 48 h, 52% loss of activity
-
0C, storage medium, no loss of activity after several hours, 30% loss of activity after 12 h, 80% loss of activity after 40 h
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DEAE-Sepharose column chromatography, Q-Sepharose column chromatography, and Sephacryl S-300 gel filtration
-
Ni-NTA agarose column chromatography
-
Ni-NTA agarose column chromatography and Sephadex G-100 gel filtration
Ni-NTA column chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli BL21(DE3)pLysS cells
His-tagged enzyme is expressed in Escherichia coli BL21(DE3) cells
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
enzyme expression can be induced by 4-nitrophenol but not 4-nitrocatechol
-