Information on EC 1.7.1.4 - nitrite reductase [NAD(P)H]

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
1.7.1.4
-
RECOMMENDED NAME
GeneOntology No.
nitrite reductase [NAD(P)H]
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
NH3 + 3 NAD(P)+ + 2 H2O = nitrite + 3 NAD(P)H + 5 H+
show the reaction diagram
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-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
alkylnitronates degradation
-
-
nitrate reduction V (assimilatory)
-
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Nitrogen metabolism
-
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non-pathway related
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SYSTEMATIC NAME
IUBMB Comments
ammonia:NAD(P)+ oxidoreductase
An iron-sulfur flavoprotein (FAD) containing siroheme. The enzymes from the fungi Neurospora crassa [1], Emericella nidulans [2] and Candida nitratophila [4] and the bacterium Aliivibrio fischeri [3] can use either NADPH or NADH as electron donor. cf. EC 1.7.1.15, nitrite reductase (NADH).
CAS REGISTRY NUMBER
COMMENTARY hide
9029-29-2
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
Lupinus, strain USDA 3045
-
-
Manually annotated by BRENDA team
cultivar RH-30
-
-
Manually annotated by BRENDA team
strain NCTC 11168
-
-
Manually annotated by BRENDA team
strain NCTC 11168
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain Bn
-
-
Manually annotated by BRENDA team
strain OR75Ch15
-
-
Manually annotated by BRENDA team
strain W3110
-
-
Manually annotated by BRENDA team
nir1; symbiotic basidomycete, 3 wild-type haploid strains, progenies of the HC1 dikaryotic strain GCA6, gene nar1
SwissProt
Manually annotated by BRENDA team
strain 128
-
-
Manually annotated by BRENDA team
strain 128
-
-
Manually annotated by BRENDA team
strains 40KI, B6, En13, NpAV and AF
-
-
Manually annotated by BRENDA team
strain CFN42
-
-
Manually annotated by BRENDA team
strain CFN42
-
-
Manually annotated by BRENDA team
Torulopsis nitratophila
-
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
NAD(P)H + NH2OH
NAD(P)+ + NH3 + H2O
show the reaction diagram
NAD(P)H + nitrite + H+
NAD(P)+ + NH3 + H2O
show the reaction diagram
nitrite + reduced benzyl viologen
ammonium hydroxide + H2O + benzyl viologen
show the reaction diagram
nitrite + reduced methyl viologen
NH3 + oxidized methyl viologen + H2O
show the reaction diagram
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
NAD(P)H + nitrite + H+
NAD(P)+ + NH3 + H2O
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
benzyl viologen
-
-
flavin
FMN
-
0.01 mM, approx. 2fold increase in activity
NADPH
additional information
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cu2+
-
depressed activity in Cu-deficient felt extracts, stimulates activity in extracts from Cu-deficient felts, depresses activity in extracts from control felts
Fe3+
-
stimulates activity in Fe-deficient felt extracts
Magnesium
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depressed activity in Mg-deficient felt extracts
Zinc
-
depressed activity in Zn-deficient felt extracts
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
1-cyclohexyl-3-(2-morpholinoethyl)-carbodiimide metho-p-toluenesulfonate
-
preincubation with NO2- completely protects
2,2'-bipyridine
2-heptyl-4-hydroxyquinoline N-oxide
-
-
8-hydroxyquinoline
antimycin A
-
0.2 mM, 45% inhibition
arsenite
Atabrine
-
-
Carbamoyl phosphate
-
-
dinitrophenol
-
-
Diquinolyl
-
-
-
Hydrazine sulfate
-
complete inhibition
hydroxylamine
Iodine
-
-
iodoacetate
-
64% inhibition
Iodosobenzoate
-
-
KCNS
-
0.1 mM, 90% inhibition of NH2OH reductase activity, nitrite reductase activity is not affected
N-ethylmaleimide
NAD(P)H
NADP+
-
0.06 mM, 13% inhibition, 0.27 mM, 43% inhibition
naphthoquinone
-
-
NH2OH
-
competitive vs. NO2-
NH4+
-
inhibits under aerobic and anaerobic conditions
nitrite
O2
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enzyme activity under anaerobic conditions is 2fold higher than under aerobic conditions
p-chloromercuribenzoate
p-hydroxymercuribenzoate
Rose bengal
-
preincubation with NO2- completely protects
rotenone
-
0.1 mM, 56% inhibition
salicylic acid
-
-
SO32-
Sodium diethyldithiocarbamate
-
-
sodium disulfite
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Thiourea
-
-
additional information
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
nitrate
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2fold enhanced induction independently of the NH4+ concentration, growth stimulation is higher on nitrate than on nitrite
pyridoxine
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stimulates activity in crude extracts from pyridoxine-deficient felts, no activation of purified enzyme
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00002 - 0.05
FAD
0.05
FMN
-
-
2.5 - 5.3
hydroxylamine
0.0063 - 0.15
NADH
0.015
NADPH
-
-
0.019
nitrite
Torulopsis nitratophila
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-
0.0048 - 0.8
NO2-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
130
NO2-
Neurospora crassa
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-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2
1,10-phenanthroline
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-
6
hydroxylamine
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0.005
KCN
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24
NH2OH
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-
0.52
NO2-
-
-
0.002
p-hydroxymercuribenzoate
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2
sodium disulfite
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5
Sodium sulfide
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-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0000205
-
after 96 h incubation under anaerobic conditions, in a medium amended with nitrite
0.000508
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after 96 h incubation under anaerobic conditions, in a medium amended without nitrite
0.06
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activity under aerobic conditions in presence of NH4+
0.065
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activity in cells grown anaerobically in the presence of 20 mM NO2-
0.079
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activity in cells grown anaerobically in the presence of 20 mM NO3-
0.1
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activity under aerobic conditions
0.13
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activity under anaerobic conditions in presence of NH4+
0.35
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activity in cytosolic extracts
0.422
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reduction of NO2-
0.504
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activity in cytosolic extracts from cells grown anaerobically with 1 mM nitrite
0.52
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NADH + NO2-
0.73
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NADPH + NO2-
0.868
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reduction of hydroxylamine
14.6
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activity in extracts from aerobic grown cells
33.4
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activity in extracts from anaerobic grown cells
1800
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activity in crude extracts of cells grown in the presence of 50 mM urea
5900
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activity in crude extracts of cells grown in the presence of 50 mM glutamine
17300
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activity in crude extracts of cells grown in the presence of 50 mM ammonium sulfate
24000
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activity in crude extracts of cells grown in the presence of 50 mM asparagine
60000
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activity in crude extracts of cells grown in the presence of 50 mM potassium nitrate
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5
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assay at
7.5 - 8.5
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 11
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pH 4.0: 40% activity, pH 11.0: 70% activity
6.5 - 9.5
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pH 6.5: about 50% activity, pH 9.5: about 15% activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
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assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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free-living cells, yeast extract/mannitol medium
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Escherichia coli (strain K12)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Pectobacterium atrosepticum (strain SCRI 1043 / ATCC BAA-672)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
67000
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sucrose density gradient centrifugation
88000
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2 * 88000, SDS-PAGE
140000
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2 * 140000, SDS-PAGE
190000
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glycerol density gradient centrifugation
289400
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deduced from amino acid composition
290000
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gel filtration, sucrose density gradient centrifugation
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40
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80% activity is lost after 4 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
dialysis against water or a variety of buffers deactivates
-
dialysis for 15 h against 1 mM cyanide, 1 mM 8-hydroxyquinoline or 1 mM EDTA followed by dialysis against 0.05 M phosphate buffer, no decrease in activity
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-17°C, overnight, 50% loss of activity
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate, DEAE-cellulose, DEAE-Sephadex
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
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expression in Escherichia coli as lacZ transcriptional fusion
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gene nir1, DNA and amino acid sequence determination and analysis, gene nir1 is part of the nitrate assimilation gene cluster together with genes encoding a high-affinity nitrate transporter and a nitrate reductase, the intergenic region could also act as bidirectional promotor, organisation of coding sequences
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
I289A
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mutant with decrease in NiR activity
I289V
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mutant with slight increase in NiR activity
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
-
microplate high-throughput nitrite reductase assay
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