Information on EC 1.14.13.167 - 4-nitrophenol 4-monooxygenase

for references in articles please use BRENDA:EC1.14.13.167
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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.13.167
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RECOMMENDED NAME
GeneOntology No.
4-nitrophenol 4-monooxygenase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
4-nitrophenol + NADPH + H+ + O2 = 1,4-benzoquinone + nitrite + NADP+ + H2O
show the reaction diagram
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
4-nitrophenol degradation I
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Aminobenzoate degradation
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Microbial metabolism in diverse environments
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SYSTEMATIC NAME
IUBMB Comments
4-nitrophenol,NAD(P)H:oxygen 4-oxidoreductase (4-hydroxylating, nitrite-forming)
Contains FAD. The enzyme catalyses the first step in a degradation pathway for 4-nitrophenol, the oxidation of 4-nitrophenol at position 4 with the concomitant removal of the nitro group as nitrite. The enzyme from the bacterium Pseudomonas sp. strain WBC-3 also catalyses EC 1.14.13.166, 4-nitrocatechol 4-monooxygenase.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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UniProt
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2,4,6-trinitrophenol + NADH + H+ + O2
?
show the reaction diagram
2-chloro-4-nitrophenol + NADPH + H+ + O2
2-chloro-1,4-benzoquinone + nitrite + NADP+ + H2O
show the reaction diagram
3-methyl-4-nitrophenol + NADPH + H+ + O2
3-methyl-1,4-benzoquinone + nitrite + NADP+ + H2O
show the reaction diagram
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?
4-nitrocatechol + NADH + H+ + O2
1,2,4-trihydroxybenzene + nitrite + NAD+ + H2O
show the reaction diagram
4-nitrocatechol + NADPH + H+ + O2
?
show the reaction diagram
lower activity compared to 4-nitrophenol
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?
4-nitrophenol + NADH + H+ + O2
1,4-benzoquinone + nitrite + NAD+ + H2O
show the reaction diagram
4-nitrophenol + NADH + H+ + O2
?
show the reaction diagram
4-nitrophenol + NADPH + H+ + O2
1,4-benzoquinone + nitrite + NADP+ + H2O
show the reaction diagram
4-nitrophenol + NADPH + H+ + O2
4-benzoquinone + nitrite + 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
2-chloro-4-nitrophenol + NADPH + H+ + O2
2-chloro-1,4-benzoquinone + nitrite + NADP+ + H2O
show the reaction diagram
3-methyl-4-nitrophenol + NADPH + H+ + O2
3-methyl-1,4-benzoquinone + nitrite + NADP+ + H2O
show the reaction diagram
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?
4-nitrophenol + NADPH + H+ + O2
1,4-benzoquinone + nitrite + NADP+ + H2O
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADPH
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
addition of Mg2+ or Mn2+ ions has no effect on the activity
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0062
2-chloro-4-nitrophenol
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pH 7.4, 30C, recombinant enzyme
0.0203
3-Methyl-4-nitrophenol
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pH and temperature not specified in the publication
0.012 - 0.0254
4-nitrophenol
0.1374
NADPH
His-tagged enzyme, in 20 mM phosphate buffer (pH 7.0), temperature not specified in the publication
additional information
additional information
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0000046
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using 2,4,6-trinitrocatechol as substrate, pH and temperature not specified in the publication
0.000025
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using 4-nitrocatechol as substrate, pH and temperature not specified in the publication
0.0000264
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using 4-nitrophenol as substrate, pH and temperature not specified in the publication
0.4
crude extract, at pH 7.0 and 30C
1.73
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mutant strain SJ98DELTApnpA1 cell extract, pH 7.5, 30C, substrate 2-chloro-4-nitrophenol
2.1
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cell extract, pH 7.4, 30C, substrate 2-chloro-4-nitrophenol
3.74
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mutant strain SJ98DELTApnpA1 cell extract, pH 7.5, 30C, substrate 4-nitrophenol
4.8
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cell extract, pH 7.4, 30C, substrate 4-nitrophenol
5.6
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pH 7.4, 30C, substrate 4-nitrophenol
6.8
purified enzyme, at pH 7.0 and 30C
7.2
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pH 7.4, 30C, substrate 2-chloro-4-nitrophenol
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.4
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assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
46600
x * 46600, calculated from amino acid sequence
47000
1 * 47000, His-tagged enzyme, gel filtration
59700
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x * 59700, 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, 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
Ni-NTA agarose column chromatography
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Ni-NTA agarose column chromatography and Sephadex G-100 gel filtration
Ni-NTA column chromatography
recombinant N-terminally His6-tagged enzyme PnpA from Escherichia coli strain Rosetta(DE3)pLysS by nickel affinity chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli Rosetta(DE3)pLysS cells
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gene pnpA, expression of the pnp cluster is detected only under inducing conditions, and pnpABA1CDEFG is a single transcriptional operon, genetic structure, RT-PCR and real-time PCR enzyme expression analysis, recombinant expression of N-terminally His6-tagged enzyme PnpA in Escherichia coli strain Rosetta(DE3)pLysS
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gene pnpA1, recombinant expression of N-terminally His6-tagged enzyme PnpA in Escherichia coli strain Rosetta(DE3)pLysS
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His-tagged enzyme is expressed in Escherichia coli BL21(DE3) cells
the enzyme is encoded in the pnpABA1CDEF cluster, which is involved in para-nitrophenol (PNP) and 2-chloro-4-nitrophenol (2C4NP) catabolism and is also likely responsible for 3-methyl-4-nitrophenol (3M4NP) degradation in strain SJ98, real-time quantitative PCR enzyme expression analysis, recombinant expression of N-terminally His6-tagged enzyme PnpA in Escherichia coli strain Rosetta(DE3)pLysS
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
0.0018 mg/ml of para-benzoquinone reductase PnpB enhances the 4-nitrophenol degradation activity by para-nitrophenol 4-monooxygenase by about 2fold
2-chloro-4-nitrophenol degradation is induced by either 2-chloro-4-nitrophenol or 4-nitrophenol, negligible activity in uninduced cells
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enzyme expression can be induced by 4-nitrophenol but not 4-nitrocatechol
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information