Information on EC 1.7.1.15 - nitrite reductase (NADH)

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The enzyme appears in viruses and cellular organisms

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EC NUMBER
COMMENTARY hide
1.7.1.15
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RECOMMENDED NAME
GeneOntology No.
nitrite reductase (NADH)
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
NH3 + 3 NAD+ + 2 H2O = nitrite + 3 NADH + 5 H+
show the reaction diagram
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Nitrogen metabolism
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Microbial metabolism in diverse environments
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SYSTEMATIC NAME
IUBMB Comments
ammonia:NAD+ oxidoreductase
An iron-sulfur flavoprotein (FAD) containing siroheme. This prokaryotic enzyme is specific for NADH. In addition to catalysing the 6-electron reduction of nitrite to ammonia, the enzyme from Escherichia coli can also catalyse the 2-electron reduction of hydroxylamine to ammonia. cf. EC 1.7.1.4, nitrite reductase [NAD(P)H].
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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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 ferricytochrome c + NADH
2 ferrocytochrome c + NAD+ + H+
show the reaction diagram
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?
horse heart cytochrome c + NADH + H+
?
show the reaction diagram
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?
hydroxylamine + NADH + H+
?
show the reaction diagram
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?
K3Fe(CN)6 + NADH + H+
?
show the reaction diagram
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?
NADH + H+ + oxidized 2,6-dichlorophenolindophenol
NAD+ + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
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?
nitrite + NADH + H+
ammonia + NAD+ + H2O
show the reaction diagram
nitrite + reduced benzyl viologen
ammonia + oxidized benzyl viologen
show the reaction diagram
nitrite + reduced methyl viologen
ammonia + oxidized methyl viologen
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
nitrite + NADH + H+
ammonia + NAD+ + H2O
show the reaction diagram
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Fe-S center
siroheme
additional information
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the enzyme does not contain FMN
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Iron
contains 2.8 mol iron per mol of protein
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,2'-dipyridyl
68% inhibition at 2 mM
azide
36% inhibition at 2 mM
Cyanate
27% inhibition at 2 mM
cyanide
69% inhibition at 2 mM
dithioerythritol
52% inhibition at 2 mM
KCN
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although 1 mM KCN completely inhibits hydroxylamine reduction, it does not inhibit the reduction of K3Fe(CN)6 and only decreases the rate of cytochrome c reduction by 12%
NAD+
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NAD+ shows mixed product inhibition with respect to NADH and mixed or uncompetitive inhibition with respect to hydroxylamine
nitrite
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nitrite decrease the rate of NADH-dependent reduction of cytochrome c by 77% and that of Fe(CN)63- by 90%
o-phenanthroline
72% inhibition at 2 mM
p-chloromercuribenzoate
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the reduction of cytochrome c, K3Fe(CN)6 and hydroxylamine is completely inhibited (more than 99%) by 0.02 mM p-chloromercuribenzoate
p-hydroxymercurybenzoate
48% inhibition at 2 mM
additional information
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0039
cytochrome c
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apparent value, at pH 8.0 and 30°C
2.91
hydroxylamine
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apparent value, at pH 8.0 and 30°C
0.1
K3Fe(CN)6
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apparent value, at pH 8.0 and 30°C
0.02 - 0.0488
NADH
0.011 - 0.5
nitrite
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.052
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crude extract, at pH 8.0 and 30°C
2
purified enzyme, at pH 9.0 and 30°C
62.4
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after 121fold purification, at pH 8.0 and 30°C
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
12000
x * 12000, NirD polypeptide, SDS-PAGE
86600
x * 86600, calculated from amino acid sequence
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SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
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2 * 88000, SDS-PAGE
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 30
the nitrite reductase activities of concentrated mixtures prepared on ice do not increase during subsequent incubation at 30°C or at 4°C
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
1 mM nitrite and 5O mM hydroxylamine are equally effective in decreasing the loss of activity after 24h at 40°C from 40% to 10%. No increase in catalytic activity is detected when enzyme is pre-incubated for 15 min with either of these substrates, even when NAD+ is omitted from the assay mixture
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation
DEAE-cellulose column chromatography
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Ni-NTA column chromatography
purified in the presence of 1 mM NO2- and 0.01 mM FAD by ammonium sulfate precipitation, DEAE-cellulose column chromatography, DEAE-Sephadex gel filtration, and Sephadex G-25 gel filtration
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli JCB323 cells
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expressed in Escherichia coli JM109 cells
expressed in Escherichia coli strain JCB387
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
gene expression is co-ordinately repressed by oxygen
gene expression is induced during anaerobic growth
under conditions of oxygen limitation, nasDEF expression and nitrite reductase activity are significantly induced. ResDE is mainly (if not exclusively) responsible for nasDEF expression during anaerobic growth regardless of nitrogen sources
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Show AA Sequence (180 entries)
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LINKS TO OTHER DATABASES (specific for EC-Number 1.7.1.15)
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)