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Enzyme Nomenclature
Information on EC 1.7.1.3 - nitrate reductase (NADPH)
for references in articles please use BRENDA:EC1.7.1.3
Word Map on EC 1.7.1.3
- 1.7.1.3
- neurospora
- crassa
- molybdenum
- nidulans
- aspergillus
-
nadph-cytochrome
-
positive-acting
-
molybdenum-containing
- viologen
- molybdopterin
-
pathway-specific
- tungstate
-
reductase-deficient
-
autogenous
-
nitrogen-related
-
nadh:nitrate
-
viologen-nitrate
- medicine
- synthesis
- analysis
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Specify your search results
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
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EC NUMBER 
COMMENTARY 
1.7.1.3
-
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RECOMMENDED NAME
GeneOntology No.
nitrate reductase (NADPH)
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
nitrite + NADP+ + H2O = nitrate + NADPH + H+
; An iron-sulfur molybdenum flavoprotein
-
-
-
nitrite + NADP+ + H2O = nitrate + NADPH + H+
random order rapid-equilibrium mechanism, two functional parts: 1. heat labile, FAD and haemoprotein containing, binds NADPH and transfers electrons from NADPH via FAD and perhaps cytochrome b to acceptors such as cytochrome c, 2. heat stable, molybdenum containing, accepts electrons from reduced viologen dyes and transfers them to nitrate
-
nitrite + NADP+ + H2O = nitrate + NADPH + H+
multicenter redox enzyme. Ser920, Arg921 and Arg932 are suggested to be the key enzymes to investigate for a role in determining pyridine nucleotide specificity. Arg932 may be playing a role in binding the adenine ring of NADPH
-
nitrite + NADP+ + H2O = nitrate + NADPH + H+
sulfhydryl groups may participate in the binding of the protein subunits
-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
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SYSTEMATIC NAME
IUBMB Comments
nitrite:NADP+ oxidoreductase
An iron-sulfur molybdenum flavoprotein.
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CAS REGISTRY NUMBER
COMMENTARY
9029-28-1
-
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
nar1; symbiotic basidomycete, 3 wild-type haploid strains, progenies of the HC1 dikaryotic strain GCA6, gene nar1
SwissProt
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
active site formation of eukaryotic nitrate reductase is an autonomous process intrinsically tied to nitrate reductase dimerization, molybdenum cofactor-dependent enzyme maturation, overview
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
chlorate + NADPH
chlorite + NADP+
-
lower affinity than to nitrate, appears to be toxic or its product
-
?
nitrate + NADPH
nitrite + NADP+
-
enzyme is essential for reduction of nitrate to ammonia, under oxic conditions the enzyme is involved in nitrate assimilation, under anoxic conditions the enzyme is used for dissimilatory nitrate reduction, transcription regulation mechanism via ammonium, nitrate, and O2 concentrations, overview
-
-
?
nitrate + NADPH
nitrite + NADP+
enzyme is involved in nitrate assimilation
-
-
?
nitrate + NADPH
nitrite + NADP+
enzyme is involved in nitrate assimilation, it serves the nitrogen nutrition of the host plant of Tuber borchii in symbiosis
-
-
?
nitrate + NADPH
nitrite + NADP+
enzyme is involved in nitrate assimilation, it serves the nitrogen nutrition of the host plant of Tuber borchii in symbiosis
-
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
some mutants use hypoxanthine as nitrogen source, 4.5-S cytochrome-c reductase activity
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
4 activities: NADPH-nitrate reductase, FADH-nitrate reductase, reduced methyl viologen-nitrate reductase and NADPH-cytochrome c reductase
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
two associated activities: cytochrome c reductase and reduced viologen dye:nitrate reductase
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
catalyzes the NADPH-linked reduction of ferricyanide and 2,6-dichlorophenolindophenol, chlorate- and bromate-dependent NADPH oxidation, and FMNH-linked nitrate reduction
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
some mutants use hypoxanthine as nitrogen source, 4.5-S cytochrome-c reductase activity
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
associated cytochrome c reductase activity
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
key enzyme in the assimilation path of nitrate to ammonium. Crude extracts possess endogenous NADPH regenerating systems capable of providing reducing equivalents for effective nitrate reduction in vitro
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
20-fold higher activity with NADPH than with NADH
-
ir
nitrate + NADPH
nitrite + NADP+ + H2O
-
4 activities: NADPH-nitrate reductase, FADH-nitrate reductase, reduced methyl viologen-nitrate reductase and NADPH-cytochrome c reductase
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
4 activities: NADPH-nitrate reductase, FADH-nitrate reductase, reduced methyl viologen-nitrate reductase and NADPH-cytochrome c reductase
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
4 activities: NADPH-nitrate reductase, FADH-nitrate reductase, reduced methyl viologen-nitrate reductase and NADPH-cytochrome c reductase
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
associated FAD-nitrate reductase and methylviologen-nitrate activity
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
first step in nitrate assimilation
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
key enzyme in the assimilation path of nitrate to ammonium. Crude extracts possess endogenous NADPH regenerating systems capable of providing reducing equivalents for effective nitrate reduction in vitro
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
first step in the reduction of nitrate to ammonia, biosynthesis of amino acids and other nitrogen-containing cell constituents
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
20-fold higher activity with NADPH than with NADH
-
ir
nitrate + NADPH
nitrite + NADP+ + H2O
-
4 activities: NADPH-nitrate reductase, FADH-nitrate reductase, reduced methyl viologen-nitrate reductase and NADPH-cytochrome c reductase
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
first step in the reduction of nitrate to ammonia, biosynthesis of amino acids and other nitrogen-containing cell constituents
-
?
nitrate + reduced methyl viologen
nitrite + oxidized methyl viologen
-
-
-
-
?
nitrate + reduced methyl viologen
nitrite + oxidized methyl viologen
-
-
-
-
?
additional information
?
-
enzyme expression is regulated by different inorganic and organic nitrogen sources, e.g. nitrate, ammonium, urea and glutamate, regulatory mechanism of nitrate aquisition in ectomycchorizae, overview
-
-
-
additional information
?
-
-
NADPH-dependent cytochrome c reducing activity by the holo-enzyme is determined with FAD and NADPH spectroscopically at 550 nm and 340 nm. Apo-nitrate reductase has a marginally lower, about 10% reduced cytochrome c reducing activity, which correlates to its 15% reduced heme content
-
-
-
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
additional information
?
-
Q9UVH6
enzyme expression is regulated by different inorganic and organic nitrogen sources, e.g. nitrate, ammonium, urea and glutamate, regulatory mechanism of nitrate aquisition in ectomycchorizae, overview
-
-
-
nitrate + NADPH
nitrite + NADP+
-
enzyme is essential for reduction of nitrate to ammonia, under oxic conditions the enzyme is involved in nitrate assimilation, under anoxic conditions the enzyme is used for dissimilatory nitrate reduction, transcription regulation mechanism via ammonium, nitrate, and O2 concentrations, overview
-
-
?
nitrate + NADPH
nitrite + NADP+
Q8H1T7
enzyme is involved in nitrate assimilation
-
-
?
nitrate + NADPH
nitrite + NADP+
Q8J259
enzyme is involved in nitrate assimilation, it serves the nitrogen nutrition of the host plant of Tuber borchii in symbiosis
-
-
?
nitrate + NADPH
nitrite + NADP+
Q8J259
enzyme is involved in nitrate assimilation, it serves the nitrogen nutrition of the host plant of Tuber borchii in symbiosis
-
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
key enzyme in the assimilation path of nitrate to ammonium. Crude extracts possess endogenous NADPH regenerating systems capable of providing reducing equivalents for effective nitrate reduction in vitro
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
first step in nitrate assimilation
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
key enzyme in the assimilation path of nitrate to ammonium. Crude extracts possess endogenous NADPH regenerating systems capable of providing reducing equivalents for effective nitrate reduction in vitro
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
first step in the reduction of nitrate to ammonia, biosynthesis of amino acids and other nitrogen-containing cell constituents
-
?
nitrate + NADPH
nitrite + NADP+ + H2O
-
first step in the reduction of nitrate to ammonia, biosynthesis of amino acids and other nitrogen-containing cell constituents
-
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
molybdopterin
-
i.e Moco/MPT, binding of molybdenum cofactor to apo-nitrate reductase is independent from other prosthetic groups, molybdenum cofactor-dependent enzyme maturation, overview. Reconstitution of Moco-free nitrate reductase with various amounts of purified Moco carrier protein
cytochrome b557
-
involved in intracellular electron transport from NADPH to nitrate
-
FAD
-
contains 5.91-7.78 nmol FAD per mg of protein, activation of NADPH-nitrate reductase activity; molybdoflavoprotein
FAD
-
indispensible role in nitrate reductase complex; molybdoflavoprotein
FAD
involved in electron transfer from NADPH to the enzyme molybdenum center where reduction of nitrate to nitrite takes place
heme
involved in electron transfer from NADPH to the enzyme molybdenum center where reduction of nitrate to nitrite takes place
molybdenum cofactor
cofactor is necessary and sufficient to induce dimer formation. The molybdenum center of nitrate reductase reconstituted in vitro from apo-enzyme and cofactor shows an EPR spectrum identical to holo-enzyme. Insertion of this cofactor into the enzyme occurs independent from the insertion of any other NR redox cofactor
molybdenum cofactor
-
i.e Moco/MPT, binding of molybdenum cofactor to apo-nitrate reductase is independent from other prosthetic groups, molybdenum cofactor-dependent enzyme maturation, overview. Reconstitution of Moco-free nitrate reductase with various amounts of purified Moco carrier protein
additional information
enzyme amino acid sequence contains a molybdate-cofactor, a cytochrome b5 heme, and a FAD binding domain
-
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Molybdenum
NH4+
-
represses the intracellular NO3- assimilation, which is required for the induction and maintenace of NADPH nitrate reductase, enzyme is regulated by NH4+ at the transcriptional level
Molybdenum
-
1 mol per mol enzyme protein, activation of enzyme after urea treatment, little stimulation
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
NADP+
-
inhibits NADPH oxidation, non-competitive with respect to nitrate
nitrite
-
competitive with respect to nitrate and non-competitive to NADPH
Phenylglyoxal
-
in 0.1 M phosphate, pH 7.3, 4 mM, inactivation after 15 min to 40% and to 20% after 60 min
p-chloromercuribenzoate
-
reversion of the inhibition by addition of reduced glutathione
additional information
-
cysteine or dithiothreitol relieve or prevent inhibition. Not inhibited by urea, glutamic acid, aspartic acid and ammonia at 10 mM
-
additional information
-
exogenous ammonium represses enzyme expression under oxic conditions in presence or absence of nitrate, while under anoxic conditions the enzyme is expressed even in the presence of ammonium, promotor activity study
-
additional information
-
intracellularly accumulated nitrite can inhibit nitrate uptake, addition of nitrite at concentrations above 5 mM is toxic and causes growth retardation
-
additional information
the nitrogen sources nitrate, glutamate, and urea induce enzyme expression, but ammonium represses it
-
additional information
-
protection of inactivation by FAD and restorage by dithiothreitol
-
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
FAD
-
activates 10fold by addition of 0.004 mM, stabilizes against heat inactivation
additional information
-
exogenous nitrate induces the enzyme expression, can be repressed by ammonium under aerobic conditions, but not under anaerobic conditions, promotor activity study
-
additional information
-
growth stimulation is higher on nitrate than on nitrite
-
additional information
the nitrogen sources nitrate, glutamate, and urea induce enzyme expression, but ammonium represses it
-
additional information
-
enzyme can be reactivated by molybdenum and dithioerythritol
-
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
Michaelis-Menten kinetics, holo-enzyme
-
0.25
nitrate
-
recombinant holo-enzyme, pH 7.5, temperature not specified in the publication
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.04 - 0.048
NADP+
-
NADPH as substrate in saturating and non-saturating concentrations, competitive
0.065
NADP+
-
nitrate as substrate in non-saturating concentrations, non-competitive
0.15
nitrite
-
NADPH as substrate in non-saturation concentration, non-competitive
0.18
nitrite
-
nitrate as substrate in saturating and non-saturating concentrations, competitive
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.019
-
wild-type enzyme, anaerobic conditions, presence of nitrate and ammonium
1.24
4.32
-
purified recombinant holo-enzyme, substrate nitrate, pH 7.5, temperature not specified in the publication
additional information
additional information
-
one unit is defined as that amount of enzyme which results in the formation of 0.000001 mM of nitrite
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.2
7.4
7.5
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25
28
30
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
additional information
additional information
the enzymatic activity is increased in root ectomycchorizae in symbiosis with the fungus Tuber borchii, while the expression of the endogenous enzyme is decreased
additional information
the enzyme expression and activity is increased in ectomycchorizae in symbiosis with Tilia platyphyllos
additional information
-
the enzyme expression and activity is increased in ectomycchorizae in symbiosis with Tilia platyphyllos
-
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PDB
SCOP
CATH
UNIPROT
ORGANISM
Pichia angusta;
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
38000
-
2 * 59000 + 2 * 38000, SDS-PAGE, after heat-treatment all polypeptide chains are 59 kDa
59000
-
2 * 59000 + 2 * 38000, SDS-PAGE, after heat-treatment all polypeptide chains are 59 kDa
91000
-
2 * 91000, homodimer, SDS-PAGE, smaller bands are caused by proteolytic cleavage
115000
-
1 * 130000 + 1 * 115000, SDS-PAGE, homodimer of 2 * 150000 suggested
130000
-
1 * 130000 + 1 * 115000, SDS-PAGE, homodimer of 2 * 150000 suggested
150000
-
1 * 130000 + 1 * 115000, SDS-PAGE, homodimer of 2 * 150000 suggested
180000
204000
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SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
heterotrimer
homodimer
multimer
tetramer
additional information
dimer
-
2 * 91000, homodimer, SDS-PAGE, smaller bands are caused by proteolytic cleavage
dimer
-
1 * 130000 + 1 * 115000, SDS-PAGE, homodimer of 2 * 150000 suggested
dimer
in presence of molybdenum cofactor, enzyme forms a dimer, gel filtration, sedimentation velocity analysis
dimer
-
1 * 130000 + 1 * 115000, SDS-PAGE, homodimer of 2 * 150000 suggested
-
homodimer
-
holo-enzyme, determination with SDS-PAGE, gel filtration and analytical ultracentrifugation. The enzyme has a largely hydrophobic dimer interface
tetramer
-
2 * 59000 + 2 * 38000, SDS-PAGE, after heat-treatment all polypeptide chains are 59 kDa
additional information
-
nit-1 mutant enzyme is the apoprotein of nitrate reductase
additional information
-
aggregate of two different polypeptide chains: one responsible for transport of electrons fom NADPH to FAD or cytochrome c, nit-1 enzyme. The other transfers electrons from FAD via molybdenum to nitrate, nit-3 enzyme; nit-1 mutant enzyme is the apoprotein of nitrate reductase
additional information
-
the apo-enzyme dissociates completely into monomers, the ratio between monomeric and dimeric apo-NR does not change significantly upon a 20fold dilution. Active site formation of eukaryotic nitrate reductase is an autonomous process intrinsically tied to nitrate reductase dimerization, molybdenum cofactor-dependent enzyme maturation, overview. Enzyme domain structure, overview
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Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
vapor diffusion against a reservoir of solution 7 from the Hampton Crystal Screen II
-
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5 - 9
-
purified native nitrate reductase, maintaining 50% of its original activity
741637
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
10
20 - 60
-
purified native nitrate reductase, pH 7.2, maintaining 50% of its original activity
25
35
-
wild-type half-life: 18.8 min, various temperature sensitive mutant strains: 3.8-18.7 min
37
40
37
-
absence of FAD, rapid loss of activity, decreases to 50% in less than 20 min
40
-
less than 50% loss of activity in 60 min for wild type and S920D mutant
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
glutathione stabilizes the enzyme at 1 mM, cysteine as well but not as effective
-
loss of activity with ammonium chloride or ammonium nitrate at 10 mM to growing cells, faster decay in washed and resuspended cells
-
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ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Ethanol
Glycerol
enzyme activity is strongly negatively impacted by increase in solution viscosity using 50% glycerol
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-10°C, 0.1 M sodium phosphate buffer, pH 7.3, 0.17 M NaCl, 1 mM dithiothreitol, 5 m M EDTA, 0.5 mM PMSF, 1%ethanol, 30% glycerol
-
-10°C, 50 mM sodium phosphate, pH 6.9, 30% glycerol, 0.5 mM EDTA, 0.5 mM dithiothreitol, 0.5 mM phenylmethylsulfonylfluoride, 0.1 mM FAD, stable for at least 6 months
-
-15°C, fraction II, most stable, optimal pH: 7.0, 10-20% loss of activity after a month
-
-15°C, fraction V, quite unstable, overnight, loses at least half of its activity
-
-80°C, 0.02 mM FAD, several months with only slight loss of the activity
-
5°C, urea-treated, 0.1 M potassium phosphate buffer, pH 7.3, 5 mM EDTA, 2 mM dithioerythritol, 0.5 mg/ml or more bovine serum albumin, stable for 18 h without loss of activity
-
half life of about 2 h with ammonium sulfate or ammonium chloride as sole N-source
-
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium slufate precipitation, ion-exchange, gel filtration, hydroxylapatite column. FAD and EDTA essential in all buffers during purification
-
immobilized metal affinity chromatography and SourceQ15 column anion exchange chromatography
-
metal-chelate affinity chromatography for his-tagged proteins, ammonium sulfate fractionation, affinity chromatography
-
native enzyme 70fold by ultrafiltration, anion exchange chromatography, and gel filtration
-
partially purification of wild-type and nit-3 mutant by streptomycin sulfate precipitation and ammonium sulfate fractionation, of nit-1 mutant by protamine sulfate fractionation, ammonium sulfate precipitation and gel filtration
-
protamine sulfate and ammonium sulfate precipitation, hemoglobin-Sepharose column, Bio-Gel A, FAD-Sepharose affinity column. proteolysis during extensive purification
-
quick purification by immunoprecipitation with monospecific anti-nitrate reductase serum, stabilization during purification and protection of proteolytic cleavage by addition of phenylmethylsulphonyl fluoride
-
recombinant enzyme from Escherichia coli strains TP1000, RK5206, and RK5204 by a two-step affinity purification
-
streptomycin sulfate and ammonium sulfate precipitation, ion-exchange, gel filtration, isoelectric focusing, FAD-affinity
-
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
construction of a fusion gene comprising gene the promotor of gene niaD and the Escherichia coli beta-glucuronidase gene, i.e. GUS, for analysis of promotor activity in Aspergillus nidulans
-
gene nar1, DNA and amino acid sequence determination and analysis, expression study, gene nar1 is part of the nitrate assimilation gene cluster together with genes encoding a high-affinity nitrate transporter and a nitrite reductase, the intergenic region could also act as bidirectional promotor, organisation of coding sequences
gene tbnr1, DNA and amino acid sequence determination and analysis, expression analysis
gene tpnr1, DNA sequence determination and analysis, construction of a cDNA library
sequence comparisons, recombinant enzyme expression of wild-type and mutant enzymes in Escherichia coli strains TP1000, RK5206, and RK5204
-
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
H654A/H677A
-
site-directed mutagenesis, CD spectroscopy shows no negative effects of the introduced mutations on protein secondary structure in comparison to the wild-type protein, but the mutant contains no heme, while the FAD binding ability is not significantly disturbed
R778E
-
site-directed mutagenesis, an FAD-binding mutant. The mutant binds essentially the same amount of Moco as does the wild type protein
R921S
-
little impact on NADPH and NADH activity, no importance for pyridine nucleotide specificity
R921T
-
little impact on NADPH and NADH activity, no importance for pyridine nucleotide specificity
R932Q
-
1/4 wild type NADPH activity is retained, twice as much NADH activity is present as compared to wild type
R932S
-
1/10 wild type NADPH activity is retained, 2/3 of wild type NADH activity
S920D
-
important for the enzyme's interaction with the pyridine nucleotide substrates. Mutant retains ~2% of the NADPH activity of the wild type while it has an increased NADH activity, ~15% higher. It is concluded that Ser920 is a ligand involved in binding the 2' phosphate of NADPH in the wild type enzyme
S920D/R932S
-
greatest decrease in NADPH activity of all created mutants, shows that Arg932 is a residue interacting with the pyridine nucleotide coenzyme electron donors and that Ser920 and Arg932 have effects on substrate binding and catalytic activity. Both residues may be ligands to the 2' phosphate of NADPH in the wild type cyt b reductase fragment of nitrate reductase
additional information
additional information
-
studies of temperature-sensitive mutations, niaD gene: mutation leads to loss of a 4.5-S cytochrome-c reductase activity, which is a subunit of nitrate reductase. It is suggested that neither the product of the cnxE nor the cnyF genes form part of the nitrate reductase molecule, but some catalytic role in cofactor formation, niaD and cnxH seem to be structural genes
additional information
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different structural gene (niaD) and cofactor gene (cnx) mutants are analyzed concerning their flavin and molybdenum content
additional information
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studies of temperature-sensitive mutations, niaD gene: mutation leads to loss of a 4.5-S cytochrome-c reductase activity, which is a subunit of nitrate reductase. It is suggested that neither the product of the cnxE nor the cnyF genes form part of the nitrate reductase molecule, but some catalytic role in cofactor formation, niaD and cnxH seem to be structural genes
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additional information
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the purified native enzyme is successfully used for synthesis of silver nanoparticles in an NADPH-dependent manner using gelatin as a capping agent, analysis by X-ray diffraction, dynamic light scattering spectroscopy, and transmission and scanning electron microscopy, overview. 0.2 M Phosphate buffer, pH 7.2, 1 mM silver nitrate as the enzyme substrate, 0.1 mg gelatin as a capping agent, 1 mM 4-hydroxyquinoline as an electron carrier, 1 mM NADPH as enzyme cofactor, and 0.1 mg of purified fungal nitrate reductase are incubated at 25°C for 5 h. The stable nonaggregating nanoparticles are spherical in shape with an average size of 50 nm and a zeta potential of -34.3. The synthesized nanoparticles show a strong growth inhibitory antimicrobial activity against all tested human pathogenic fungi and bacteria, overview
additional information
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the purified native enzyme is successfully used for synthesis of silver nanoparticles in an NADPH-dependent manner using gelatin as a capping agent, analysis by X-ray diffraction, dynamic light scattering spectroscopy, and transmission and scanning electron microscopy, overview. 0.2 M Phosphate buffer, pH 7.2, 1 mM silver nitrate as the enzyme substrate, 0.1 mg gelatin as a capping agent, 1 mM 4-hydroxyquinoline as an electron carrier, 1 mM NADPH as enzyme cofactor, and 0.1 mg of purified fungal nitrate reductase are incubated at 25°C for 5 h. The stable nonaggregating nanoparticles are spherical in shape with an average size of 50 nm and a zeta potential of -34.3. The synthesized nanoparticles show a strong growth inhibitory antimicrobial activity against all tested human pathogenic fungi and bacteria, overview
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additional information
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several mutations of recombinant cyt b reductase fragment of nitrate reductase in the region Ser920, Arg921 and Arg932 are created. Conversion from NADPH-specific to virtually NADH-specific cyt b reductase fragment of nitrate reductase
additional information
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nit-1 mutant, suggested to produce the complete apoenzyme
additional information
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nit-1 mutant: lacks all activities except FAD-dependent NADPH:cytochrome c reductase activity,nit-2 mutant: reduced FAD:- and reduced methyl viologen:nitrate reductase activities but lacks the other two activities
additional information
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nit-3 mutant (FGSC 262): reduced FAD-nitrate reductase and reduced methylviologen-nitrate reductase activities
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
analysis
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sequential injection analysis flow system for determination of nitrites and nitrates in human serum
medicine
synthesis
medicine
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the purified native enzyme is successfully used for synthesis of silver nanoparticles in an NADPH-dependent manner using gelatin as a capping agent, analysis by X-ray diffraction, dynamic light scattering spectroscopy, and transmission and scanning electron microscopy, overview. The stable nonaggregating nanoparticles are spherical in shape with an average size of 50 nm and a zeta potential of -34.3. The synthesized nanoparticles show a strong growth inhibitory antimicrobial activity against all tested human pathogenic fungi and bacteria, overview
medicine
-
the purified native enzyme is successfully used for synthesis of silver nanoparticles in an NADPH-dependent manner using gelatin as a capping agent, analysis by X-ray diffraction, dynamic light scattering spectroscopy, and transmission and scanning electron microscopy, overview. The stable nonaggregating nanoparticles are spherical in shape with an average size of 50 nm and a zeta potential of -34.3. The synthesized nanoparticles show a strong growth inhibitory antimicrobial activity against all tested human pathogenic fungi and bacteria, overview
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synthesis
-
the purified native enzyme is successfully used for synthesis of silver nanoparticles in an NADPH-dependent manner using gelatin as a capping agent, analysis by X-ray diffraction, dynamic light scattering spectroscopy, and transmission and scanning electron microscopy, overview. The stable nonaggregating nanoparticles are spherical in shape with an average size of 50 nm and a zeta potential of -34.3. The synthesized nanoparticles show a strong growth inhibitory antimicrobial activity against all tested human pathogenic fungi and bacteria, overview
synthesis
-
the purified native enzyme is successfully used for synthesis of silver nanoparticles in an NADPH-dependent manner using gelatin as a capping agent, analysis by X-ray diffraction, dynamic light scattering spectroscopy, and transmission and scanning electron microscopy, overview. The stable nonaggregating nanoparticles are spherical in shape with an average size of 50 nm and a zeta potential of -34.3. The synthesized nanoparticles show a strong growth inhibitory antimicrobial activity against all tested human pathogenic fungi and bacteria, overview
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