Information on EC 1.7.1.B3 - aromatic nitroreductase [NADPH]

for references in articles please use BRENDA:EC1.7.1.B3
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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.7.1.B3
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
aromatic nitroreductase [NADPH]
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
an aromatic amine + 3 NADP+ + 2 H2O = an aromatic nitrate + 3 NADPH + 3 H+
show the reaction diagram
an aromatic amine + NADP+ + H2O = an aromatic hydroxylamine + NADPH + H+
show the reaction diagram
(1a)
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an aromatic hydroxylamine + NADP+ = an aromatic nitric oxide + NADPH + H+
show the reaction diagram
(1b)
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-
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an aromatic nitric oxide + NADP+ + H2O = an aromatic nitrate + NADPH + H+
show the reaction diagram
(1c)
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SYSTEMATIC NAME
IUBMB Comments
aromatic nitrate:NADPH oxidroreductase
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
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
2,4,6-trinitrotoluene + NADPH + H+
4-hydroxylamino-2,6-dinitrotoluene + NADP+ + H2O
show the reaction diagram
2,4,6-trinitrotoluene + NADPH + H+
? + NADP+ + H2O
show the reaction diagram
2,4-dinitrotoluene + NADPH + H+
? + NADP+ + H2O
show the reaction diagram
-
-
-
?
2-amino-4,6-dinitrotoluene + NADPH + H+
? + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
4-amino-2,6-dinitrotoluene + NADPH + H+
? + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
5-(aziridin-1-yl)-2,4-dinitrobenzamide + NADPH + H+
? + NADP+
show the reaction diagram
m-dinitrobenzene + NADPH + H+
? + NADP+
show the reaction diagram
-
-
-
-
?
methyl 4-nitrobenzoate + NADPH + H+
? + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
nifuroxime + NADPH + H+
? + NADP+
show the reaction diagram
-
-
-
-
?
nitro-CBI-5-[(dimethylamino)ethoxy]indole + NAD(P)H + H+
? + NAD(P)+
show the reaction diagram
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a nitro-CBI prodrug, low activity
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-
?
nitrobenzene + NADPH + H+
? + NADP+
show the reaction diagram
-
-
-
-
?
nitrofurantoin + NADPH + H+
? + NADP+
show the reaction diagram
-
-
-
-
?
nitrofurazone + NADPH + H+
5-(hydroxyamino)furan-2-carbaldehyde semicarbazone + NADP+ + H2O
show the reaction diagram
o-dinitrobenzene + NADPH + H+
? + NADP+
show the reaction diagram
-
-
-
-
?
p-dinitrobenzene + NADPH + H+
? + NADP+
show the reaction diagram
-
-
-
-
?
p-nitroacetophenone + NADPH + H+
? + NADP+
show the reaction diagram
-
-
-
-
?
p-nitrobenzaldehyde + NADPH + H+
? + NADP+
show the reaction diagram
-
-
-
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?
p-nitrobenzoic acid + NADPH + H+
? + NADP+
show the reaction diagram
-
-
-
-
?
tetryl + NADPH + H+
? + NADP+
show the reaction diagram
-
-
-
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?
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
2,4,6-trinitrotoluene + NADPH + H+
4-hydroxylamino-2,6-dinitrotoluene + NADP+ + H2O
show the reaction diagram
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-
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?
nitrofurazone + NADPH + H+
5-(hydroxyamino)furan-2-carbaldehyde semicarbazone + NADP+ + H2O
show the reaction diagram
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?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADPH
additional information
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NADPH is much more effective than NADH
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
dicoumarol
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inhibits Escherichia coli enzyme NfsB far more strongly than enzyme NfsA, by acting as a competitive inhibitor towards NADPH, uncompetitive towards substrate tetryl
NADP+
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inhibition of NfsA-catalyzed tetryl reduction by NADP+
additional information
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the enzyme is O2-insensitive
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.013 - 0.033
2,4,6-trinitrotoluene
0.176 - 0.366
2,4-dinitrotoluene
0.3257
2-amino-4,6-dinitrotoluene
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pH 7.0, 37C, recombinant His6-tagged enzyme
0.6427
4-amino-2,6-dinitrotoluene
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pH 7.0, 37C, recombinant His6-tagged enzyme
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0.23 - 0.487
5-(aziridin-1-yl)-2,4-dinitrobenzamide
0.046
m-dinitrobenzene
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pH 7.0, 25C
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0.00085 - 0.085
NADPH
0.031
nifuroxime
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pH 7.0, 25C
1.5
nitrobenzene
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pH 7.0, 25C
0.019
Nitrofurantoin
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pH 7.0, 25C
0.012 - 0.064
nitrofurazone
0.018
o-dinitrobenzene
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pH 7.0, 25C
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0.037
p-dinitrobenzene
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pH 7.0, 25C
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0.39
p-nitroacetophenone
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pH 7.0, 25C
0.17
p-nitrobenzaldehyde
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pH 7.0, 25C
2.5
p-nitrobenzoic acid
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pH 7.0, 25C
0.011
tetryl
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pH 7.0, 25C
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additional information
additional information
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.67 - 89
2,4,6-trinitrotoluene
0.7 - 24.94
2,4-dinitrotoluene
12.36
2-amino-4,6-dinitrotoluene
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pH 7.0, 37C, recombinant His6-tagged enzyme
1.05
4-amino-2,6-dinitrotoluene
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pH 7.0, 37C, recombinant His6-tagged enzyme
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1.08 - 36.64
5-(aziridin-1-yl)-2,4-dinitrobenzamide
55
m-dinitrobenzene
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pH 7.0, 25C
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0.46 - 16.3
NADPH
180
nifuroxime
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pH 7.0, 25C
14
nitrobenzene
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pH 7.0, 25C
136
Nitrofurantoin
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pH 7.0, 25C
0.8 - 19.37
nitrofurazone
60
o-dinitrobenzene
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pH 7.0, 25C
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27
p-dinitrobenzene
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pH 7.0, 25C
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59
p-nitroacetophenone
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pH 7.0, 25C
30
p-nitrobenzaldehyde
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pH 7.0, 25C
64
p-nitrobenzoic acid
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pH 7.0, 25C
85
tetryl
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pH 7.0, 25C
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
50 - 2697
2,4,6-trinitrotoluene
2 - 142
2,4-dinitrotoluene
38
2-amino-4,6-dinitrotoluene
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pH 7.0, 37C, recombinant His6-tagged enzyme
1.6
4-amino-2,6-dinitrotoluene
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pH 7.0, 37C, recombinant His6-tagged enzyme
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2 - 112
5-(aziridin-1-yl)-2,4-dinitrobenzamide
1195.7
m-dinitrobenzene
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pH 7.0, 25C
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5 - 191.7
NADPH
5806.5
nifuroxime
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pH 7.0, 25C
9.3
nitrobenzene
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pH 7.0, 25C
7157.9
Nitrofurantoin
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pH 7.0, 25C
10 - 1610
nitrofurazone
3333.3
o-dinitrobenzene
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pH 7.0, 25C
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729.7
p-dinitrobenzene
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pH 7.0, 25C
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151.3
p-nitroacetophenone
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pH 7.0, 25C
176.5
p-nitrobenzaldehyde
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pH 7.0, 25C
25.6
p-nitrobenzoic acid
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pH 7.0, 25C
7727.27
tetryl
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pH 7.0, 25C
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.018
dicoumarol
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pH 7.0, 25C
additional information
additional information
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inhibition kinetics
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.1
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purified recombinant mutant F46A, pH 7.0, 22C, substrate nitrofurazone
0.29
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purified recombinant mutant F46N, pH 7.0, 22C, substrate nitrofurazone
3.74
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purified recombinant mutant F46Y, pH 7.0, 22C, substrate nitrofurazone
7.75
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purified recombinant wild-type enzyme, pH 7.0, 22C, substrate nitrofurazone
52.5
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purified recombinant untagged enzyme, pH 7.0, 23C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
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assay at room temperature
23
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assay at
25
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assay at
37
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assay at
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
53000
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recombinant untagged enzyme, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
the enzyme is O2-insensitive
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742167
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme 23fold from Escherichia coli to homogeneity by dialysis, anion exchange chromatography, dialysis, two steps of Blue Sepharose affinity chromatography, and again dialysis
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recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
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recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography to over 95% purity
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
gene ipa-43d, DNA and amino acid sequence determination and analysis, genetic structure and sequence comparison, recombinant overexpression in Escherichia coli J83
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gene nfsA, recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha
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gene nfsA, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
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gene nfsA, recombinant overexpression in Escherichia coli reporter strain SOS-R2, which contains a lacZ reporter gene under control of the SOS responsive promoter sfiA, functional nitroreductase NfsA expression in human HCT-116 cells
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
F42A
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site-directed mutagenesis, the mutant enzymes shows 99% reduced activity toward nitrofurazone compared to the wild-type, and loss of helix content
F42N
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site-directed mutagenesis, the mutant enzymes shows 96% reduced activity toward nitrofurazone compared to the wild-type, and loss of helix content
F42Y
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site-directed mutagenesis, the mutant enzymes shows 52% reduced activity toward nitrofurazone compared to the wild-type
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
environmental protection
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NfsA has potential applications in the biodegradation of nitroaromatic environment pollutants, e.g. explosives
medicine
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NfsA has potential applications in the anticancer strategy gene-directed enzyme prodrug therapy