Information on EC 1.7.1.14 - nitric oxide reductase [NAD(P)+, nitrous oxide-forming]

for references in articles please use BRENDA:EC1.7.1.14
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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY hide
1.7.1.14
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RECOMMENDED NAME
GeneOntology No.
nitric oxide reductase [NAD(P)+, nitrous oxide-forming]
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
N2O + NAD(P)+ + H2O = 2 NO + NAD(P)H + H+
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
co-denitrification
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denitrificaton
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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
nitrous oxide:NAD(P) oxidoreductase
A heme-thiolate protein (P-450). The enzyme from Fusarium oxysporum utilizes only NADH, but the isozyme from Trichosporon cutaneum utilizes both NADH and NADPH. The electron transfer from NAD(P)H to heme occurs directly, not requiring flavin or other redox cofactors.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
microcosms established with soils from two representative U.S. Midwest agricultural regions that produce nitrous oxide
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
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isozyme P450nor2 acts as the electron sink under anaerobic, denitrifying conditions to regenerate NADP+ for the pentose phosphate cycle
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
DEA-NONOate + NADH + H+
N2O + NAD+ + ?
show the reaction diagram
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?
NO + N3-
N2O + ?
show the reaction diagram
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the co-denitrification reaction does not require an electron donor such as NADH
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?
NO + NAD(P)H + H+
N2O + NAD(P)+ + H2O
show the reaction diagram
NO + NADH + H+
N2O + NAD+ + H2O
show the reaction diagram
NO + NADP + H+
N2O + NADP+ + H2O
show the reaction diagram
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isozyme P450Nor2 prefers NADPH to NADH
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?
NO + NADPH + H+
N2O + NADP+ + H2O
show the reaction diagram
NO + NH4+
N2O + ?
show the reaction diagram
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the co-denitrification reaction does not require an electron donor such as NADH
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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
NO + NAD(P)H + H+
N2O + NAD(P)+ + H2O
show the reaction diagram
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?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.801
NADH
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in 50 mM sodium phosphate (pH 7.2), at 37°C
0.934
NADPH
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in 50 mM sodium phosphate (pH 7.2), at 37°C
0.02 - 0.6
NO
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
83.3 - 1200
NO
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.4
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isozyme P450nor1, isoelectric focusing
5.2
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isozyme P450nor1, isoelectric focusing
PDB
SCOP
CATH
ORGANISM
UNIPROT
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45000
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SDS-PAGE
45200
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calculated from amino acid sequence
45690
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deduced from amino acid sequence
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
P450nor in a complex with an NADH analogue, nicotinic acid adenine dinucleotide, sitting drop vapor diffusion method, using 20-30% (w/v) PEG 8000, 0.2 M sodium acetate, 0.1 M sodium cacodylate (pH 5.5-7.5)
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sitting-drop vapor diffusion method, using 36% (w/v) PEG 4000, 0.1 M MES, 10% (v/v) glycerol at pH 7.0
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, Resource Q column chromatography, Resource PHE column chromatography, and Superdex 200 gel filtration
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DEAE cellulose column chromatography, phenyl-Superose column chromatography, and Mono Q column chromatography or Resource Q column chromatography
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DEAE-cellulose column chromatography and Mono Q or Resource Q column chromatography
Ni-NTA column chromatography and Poros HQ anion-exchange column chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a 404 amino acid fragment of Nor1p is expressed in Escherichia coli JM109 cells
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expressed in Escherichia coli
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expressed in Escherichia coli JM109 (DE3) cells
expressed in Escherichia coli JM109(DE3) cells
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expression in Escherichia coli
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isozyme Cnor2 is expressed in Nicotiana tabacum BY-2 cells
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wild type and mutant enzymes are expressed in Escherichia coli JM109 cells
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
p450nor gene is present in all fungal isolates analyzed that produce N2O from NO2, whereas nirK encoding the NO-forming NO2 reductase, is amplified in only 13 to 74% of the N2O-forming isolates
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the addition of glucose as a carbon source to the culture medium leads to the production of much more isozyme P450nor2 than a non-fermentable substrate (glycerol or acetate) does
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
G76A
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the mutation decreases the NADH-dependent activity to 40% of the wild type activity
K77A
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the mutation decreases the NADH-dependent activity
Q78A
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the mutation slightly improves the NADH-dependent activity
S73A
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the mutation decreases the NADH-dependent activity
S73G/S75G
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the double mutation significantly improves NADPH-dependent Nor activity without affecting, or even slightly increasing, the NADHd-ependent activity
S73G/S75GS75G
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the mutant exhibits 79% activity of wild type with NADH
S75A
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the mutation decreases the NADH-dependent activity
T243A
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the mutant shows 3% of wild type NO reduction activity
T243N
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the mutant shows 2% of wild type NO reduction activity
T243V
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the mutant shows 0.01% of wild type NO reduction activity
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
degradation
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p450nor gene is present in all fungal isolates analyzed that produce N2O from NO2, whereas nirK encoding the NO-forming NO2 reductase, is amplified in only 13 to 74% of the N2O-forming isolates
synthesis
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coimmobilization of nitric oxide reductase and Bacillus megaterium glucose dehydrogenase for the continuous reduction of nitric oxide via cofactor recycling, using epoxide- and carboxyl-functionalized hyperporous microspheres. Enzyme activity maintenance of 158% for nitric oxide reductase and 104% for glucose dehydrogenase can be achieved