Information on EC 1.13.12.16 - nitronate monooxygenase

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

EC NUMBER
COMMENTARY hide
1.13.12.16
-
RECOMMENDED NAME
GeneOntology No.
nitronate monooxygenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ethylnitronate + O2 = acetaldehyde + nitrite + other products
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
alkylnitronates degradation
-
-
Nitrogen metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
nitronate:oxygen 2-oxidoreductase (nitrite-forming)
Previously classified as 2-nitropropane dioxygenase (EC 1.13.11.32), but it is now recognized that this was the result of the slow ionization of nitroalkanes to their nitronate (anionic) forms. The enzymes from the fungus Neurospora crassa and the yeast Williopsis saturnus var. mrakii (formerly classified as Hansenula mrakii) contain non-covalently bound FMN as the cofactor. Neither hydrogen peroxide nor superoxide were detected during enzyme turnover. Active towards linear alkyl nitronates of lengths between 2 and 6 carbon atoms and, with lower activity, towards propyl-2-nitronate. The enzyme from N. crassa can also utilize neutral nitroalkanes, but with lower activity.
CAS REGISTRY NUMBER
COMMENTARY hide
61584-55-2
-
65802-82-6
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1-hydroxybutyl-2-nitronate + O2
3-hydroxy-butane-2-one + HNO2
show the reaction diagram
-
-
-
-
?
1-hydroxybutyl-2-nitronate + O2
? + HNO2
show the reaction diagram
-
anionic, expression on the basis of the reactivity of propyl-2-nitronate with 2-nitropropane dioxygenase: 31.5
-
-
?
1-hydroxyethyl-2-nitronate + O2
glycoaldehyde + HNO2
show the reaction diagram
-
-
-
-
?
1-nitrobutane + O2
? + HNO2
show the reaction diagram
-
-
-
-
?
1-nitrobutane + O2
butyraldehyde + HNO2
show the reaction diagram
1-nitrobutane + O2
butyraldehyde + nitrite
show the reaction diagram
-
-
-
?
1-nitrohexane + O2
? + HNO2
show the reaction diagram
-
-
-
-
?
1-nitrohexane + O2
hexanaldehyde + HNO2
show the reaction diagram
1-nitrohexane + O2
hexanaldehyde + nitrite
show the reaction diagram
-
-
-
?
1-nitropentane + O2
? + HNO2
show the reaction diagram
1-nitropentane + O2
pentanaldehyde + HNO2
show the reaction diagram
1-nitropentane + O2
pentanaldehyde + nitrite
show the reaction diagram
-
-
-
?
1-nitropropane + O2
1,1-dinitropropane + HNO2
show the reaction diagram
-
-
-
-
?
1-nitropropane + O2
? + HNO2
show the reaction diagram
-
-
-
-
?
1-nitropropane + O2
propionaldehyde + HNO2
show the reaction diagram
1-nitropropane + O2
propionaldehyde + nitrite
show the reaction diagram
-
-
-
?
1-nitropropane + O2 + H2O
propionaldehyde + HNO2 + H2O2
show the reaction diagram
-
-
-
-
?
2 Cu((CH3)2CNO2)(PPh3)2 + O2
2 Cu(O2N)(PPh3)2 + 2 propan-2-one
show the reaction diagram
-
using a copper(I) aci-2-nitropropanate complex
-
-
?
2-hydroxybutyl-3-nitronate + O2
? + HNO2
show the reaction diagram
-
anionic, expression on the basis of the reactivity of propyl-2-nitronate with 2-nitropropane dioxygenase: 26.7
-
-
?
2-hydroxypentyl-3-nitronate + O2
2-hydroxy-pentane-3-one + HNO2
show the reaction diagram
-
-
-
-
?
2-hydroxypentyl-3-nitronate + O2
? + HNO2
show the reaction diagram
-
anionic, expression on the basis of the reactivity of propyl-2-nitronate with 2-nitropropane dioxygenase: 32.3
-
-
?
2-nitro-1-butanol + O2
1-hydroxy-butane-2-one + HNO2
show the reaction diagram
2-nitro-1-propanol + O2
1-hydroxy-propane-2-one + HNO2
show the reaction diagram
2-nitro-1-propanol + O2
? + HNO2
show the reaction diagram
-
-
-
-
?
2-nitro-1H-indene-1,3(2H)-dione + Cu(0) + N,N,N',N'-tetramethylethylenediamine + O2
1H-indene-1,2,3,-trione + (NO2)CuN,N,N',N'-tetramethylethylenediamine
show the reaction diagram
-
with N,N-dimethylformamid (conversion: 30%) as solvent
-
-
?
2-nitroethanol + O2
? + HNO2
show the reaction diagram
-
-
-
-
?
2-nitroethanol + O2
glycoaldehyde + HNO2
show the reaction diagram
-
13% of the activity with 2-nitropropane
-
-
?
2-nitroethanol + O2 + H2O
? + HNO2 + H2O2
show the reaction diagram
-
8.4% relative activity (1-nitropropane: 100%)
-
-
?
2-nitropropane + O2
?
show the reaction diagram
-
-
-
-
?
2-nitropropane + O2
? + HNO2
show the reaction diagram
-
-
-
-
?
2-nitropropane + O2
acetone + HNO2
show the reaction diagram
2-nitropropane + O2
acetone + nitrite
show the reaction diagram
-
-
-
?
2-nitropropane + O2 + H2O
acetone + HNO2 + H2O2
show the reaction diagram
-
96.9% relative activity (1-nitropropane: 100%)
-
-
?
3-nitro-1-butanol + O2 + H2O
? + HNO2 + H2O2
show the reaction diagram
-
15.7% relative activity (1-nitropropane: 100%)
-
-
?
3-nitro-2-butanol + O2
3-hydroxy-butane-2-one + HNO2
show the reaction diagram
3-nitro-2-butanol + O2
? + HNO2
show the reaction diagram
-
-
-
-
?
3-nitro-2-butanol + O2 + H2O
? + HNO2 + H2O2
show the reaction diagram
-
6.5% relative activity (1-nitropropane: 100%)
-
-
?
3-nitro-2-pentanol + O2
2-hydroxy-pentane-3-one + HNO2
show the reaction diagram
3-nitro-2-pentanol + O2
? + HNO2
show the reaction diagram
-
-
-
-
?
3-nitro-2-pentanol + O2 + H2O
2-hydroxy-pentane-3-one + HNO2 + H2O2
show the reaction diagram
-
116% relative activity (1-nitropropane: 100%)
-
-
?
3-nitropropionate + O2
?
show the reaction diagram
-
-
-
-
?
3-nitropropionate + O2 + H2O
? + HNO2 + H2O2
show the reaction diagram
-
0.5% relative activity (1-nitropropane: 100%)
-
-
?
3-nitropropionic acid + O2
?
show the reaction diagram
butyl-1-nitronate + O2
? + nitrite
show the reaction diagram
butyl-1-nitronate + O2
NO2- + butanal
show the reaction diagram
-
-
-
-
?
cyclohexyl nitronate + O2
? + HNO2
show the reaction diagram
-
-
-
-
?
ethyl nitronate + O2
? + nitrite
show the reaction diagram
ethyl nitronate + O2
acetaldehyde + HNO2
show the reaction diagram
-
-
-
-
?
ethylnitronate + Cu(0) + N,N,N',N'-tetramethylethylenediamine + O2
acetone + (NO2)CuN,N,N',N'-tetramethylethylenediamine
show the reaction diagram
-
with N,N-dimethylformamid (conversion: 60%) and pyridine (conversion: 90%) as solvent
-
-
?
ethylnitronate + O2
acetaldehyde + HNO2
show the reaction diagram
ethylnitronate + O2
NO2- + acetaldehyde
show the reaction diagram
-
-
-
-
?
ethylnitronate + O2 + FMNH2
acetaldehyde + nitrite + FMN + H2O
show the reaction diagram
hexyl-1-nitronate + O2
? + nitrite
show the reaction diagram
hexyl-1-nitronate + O2
NO2- + hexanal
show the reaction diagram
-
-
-
-
?
nitrocyclohexane + O2
? + HNO2
show the reaction diagram
nitrocyclohexane + O2
cyclohexanone + HNO2
show the reaction diagram
nitrocyclohexane + O2 + H2O
cyclohexanone + HNO2 + H2O2
show the reaction diagram
-
99.8% relative activity (1-nitropropane: 100%)
-
-
?
nitrocyclopentane + O2
? + HNO2
show the reaction diagram
-
-
-
-
?
nitroethane + O2
? + nitrite
show the reaction diagram
-
-
-
-
?
nitroethane + O2
acetaldehyde + HNO2
show the reaction diagram
nitroethane + O2
acetaldehyde + HNO2 + 1,1-dinitroethane
show the reaction diagram
-
in contrast with the unambiguous stoichiometry of 2-nitropropane oxidation, the nitroethane oxidation is stoichiometrically complicated; 1,1-dinitroethane and nitrate are formed as minor products
-
-
?
nitroethane + O2
acetaldehyde + nitrite
show the reaction diagram
nitroethane + O2
ethanal + nitrite
show the reaction diagram
nitroethane + O2
ethylnitronate
show the reaction diagram
-
2-nitropropane dioxygenase utilizes a branched catalytic mechanism with nitroethane as substrate. The branch point occurs at the enzyme-ethylnitronate complex and involves either the release of the nitronate or an oxidative denitrification reaction. The partitioning of the enzyme-nitronate complex results in the formation of multiple products from independent catalytic pathways with nitroethane as substrate for the enzyme. In the nonoxidative pathway, nitroethane is deprotonated by histidine 196 to generate ethylnitronate which is subsequently released from the enzyme as a reaction product. The oxidative denitrification pathway was established in previous studies of the enzyme and involves the oxidation of ethylnitronate by the enzyme bound flavin to generate acetaldehyde and nitrite as product
-
-
r
nitroethane + O2 + H2O
? + HNO2 + H2O2
show the reaction diagram
-
51.8% relative activity (1-nitropropane: 100%)
-
-
?
nitroethane + O2 + H2O
ethanal + nitrite + H2O2
show the reaction diagram
nitromethane + O2
? + HNO2
show the reaction diagram
-
-
-
-
?
nitromethane + O2
formaldehyde + HNO2
show the reaction diagram
nitromethane + O2 + H2O
? + HNO2 + H2O2
show the reaction diagram
-
7.3% relative activity (1-nitropropane: 100%)
-
-
?
pentane-1-nitronate + Cu(0) + N,N,N',N'-tetramethylethylenediamine + O2
pentaldehyde + (NO2)CuN,N,N',N'-tetramethylethylenediamine
show the reaction diagram
-
with N,N-dimethylformamid (conversion: 28%) and pyridine (conversion: 21%) as solvent
-
-
?
pentyl-1-nitronate + O2
? + HNO2
show the reaction diagram
-
-
-
-
?
pentyl-1-nitronate + O2
? + nitrite
show the reaction diagram
pentyl-1-nitronate + O2
NO2- + pentanal
show the reaction diagram
-
-
-
-
?
propionate 3-nitronate + O2
?
show the reaction diagram
-
-
-
-
?
propionate-3-nitronate + O2
?
show the reaction diagram
propyl-1-nitronate + O2
?
show the reaction diagram
-
-
-
?
propyl-1-nitronate + O2
? + nitrite
show the reaction diagram
propyl-1-nitronate + O2
NO2- + propionaldehyde
show the reaction diagram
-
-
-
-
?
propyl-1-nitronate + O2
propionaldehyde + HNO2
show the reaction diagram
-
-
-
-
?
propyl-1-nitronate + O2 + FMNH2
? + nitrite + FMN + H2O
show the reaction diagram
-
-
-
-
?
propyl-2-nitronate + Cu(0) + 1,10-phenanthroline + O2
propan-2-one + ?
show the reaction diagram
-
with methanol (conversion: 42%), MeCN (conversion: 24%), and N,N-dimethylformamid (conversion: 43%)
-
-
?
propyl-2-nitronate + Cu(0) + 2,2'-bipyridine + O2
propan-2-one + ?
show the reaction diagram
-
with methanol (conversion: 44%), MeCN (conversion: 54%), and N,N-dimethylformamid (conversion: 37%)
-
-
?
propyl-2-nitronate + Cu(0) + N,N,N',N'-tetramethylethylenediamine + O2
propan-2-one + (NO2)CuN,N,N',N'-tetramethylethylenediamine
show the reaction diagram
-
with methanol (conversion: 70%), MeCN (conversion: 49%), N,N-dimethylformamid (conversion: 71%), and pyridine (conversion: 67%) as solvent
-
-
?
propyl-2-nitronate + Cu(0) + O2
propan-2-one + ?
show the reaction diagram
-
without ligand and without solvent (conversion: 12%)
-
-
?
propyl-2-nitronate + O2
? + nitrite
show the reaction diagram
propyl-2-nitronate + O2
acetone + HNO2
show the reaction diagram
propyl-2-nitronate + O2
NO2- + acetone
show the reaction diagram
-
-
-
-
?
propyl-2-nitronate + O2 + FMNH2
? + nitrite + FMN + H2O
show the reaction diagram
propylnitronate + O2
? + HNO2
show the reaction diagram
-
anionic, expression on the basis of the reactivity of propyl-2-nitronate with 2-nitropropane dioxygenase: 57.9
-
-
?
undecan-6-nitronate + Cu(0) + N,N,N',N'-tetramethylethylenediamine + O2
undecan-6-one + (NO2)CuN,N,N',N'-tetramethylethylenediamine
show the reaction diagram
-
with N,N-dimethylformamid (conversion: 66%) and pyridine (conversion: 67%) as solvent
-
-
?
additional information
?
-
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
ethylnitronate + O2 + FMNH2
acetaldehyde + nitrite + FMN + H2O
show the reaction diagram
nitroethane + O2
acetaldehyde + nitrite
show the reaction diagram
propyl-1-nitronate + O2 + FMNH2
? + nitrite + FMN + H2O
show the reaction diagram
-
-
-
-
?
propyl-2-nitronate + O2 + FMNH2
? + nitrite + FMN + H2O
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cytochrome c
-
cytochrome c also inhibits the reaction and half-inhibition is found in the presence of 17 microM cytochrome c. The addition of cytochrome c to the reaction mixture containing 2-nitropropane and 2-nitropropane dioxygenase causes an increase in absorbance at 550 nm indicating the reduction of cytochrome c
riboflavin 5'-phosphate
-
can partially replace FAD
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Fe
-
enzyme contains 1.09 gatom of iron per mol of enzyme; enzyme contains 1.09 gatom of iron per mol of enzyme
Fe3+
-
contains 1 g atom of non-heme iron per mol of enzyme
Iron
stoichiometry of 0.02 FMN per 1 monomer of enzyme
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
3-(3,4-dihydroxyphenyl)-L-Ala
-
-
3-nitro-1-propionate
-
; competitive inhibitor, pH 7.0, 30C
4-hydroxy-2,2,6,6-tetramethyl-piperidinooxy radical
-
-
5-hydroxytryptophan
-
-
8-hydroxyquinoline
-
; strong inhibition
acetone
-
inhibits competitively against 2-nitropropane, no inhibitory effect against O2
alpha-Naphthol
-
-
cysteine
-
; marked decrease in enzyme activity
cytochrome c
dithiothreitol
EDTA
-
1 mM, relative activity remaining 95%
epinephrine
ethylnitronate
-
-
HgCl2
hydroquinone
-
-
iodoacetate
-
; 1 mM, complete inhibition
KCN
-
; strongly
m-Nitrobenzoate
-
-
N-ethylmaleimide
NADPH
-
competitive inhibitor. oxygenation is inhibited approximately 100% by 0.1 mM NADPH, respectively, although NAD+ and NADP+ are ineffective even at 1 mM. During the inhibition, NADH and NADPH are oxidized
nitrite
-
inhibits noncompetitively against 2-nitropropane, functioned as a noncompetitive inhibitor against oxygen in the presence of excess 2-nitropropane (50 mM)
Nitro blue tetrazolium
Nitromethane
-
inhibits noncompetitively
p-chloromercuribenzoate
phloroglucinol
-
-
pyrogallol
-
-
resorcinol
-
-
Superoxide dismutase
-
Thiophenol
-
-
Tiron
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,2'-dipyridyl
-
-
2,4,6-tripyridyl-triazine
-
-
-
o-phenanthroline
-
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
15 - 17.9
1-nitrobutane
1.4
1-nitrohexane
in 50 mM Tris-HCl, pH 8 at 30C
7.1
1-nitropentane
in 50 mM Tris-HCl, pH 8 at 30C
1.54 - 57
1-Nitropropane
1.61 - 33
2-Nitropropane
0.59 - 4.2
3-Nitro-2-butanol
1.04 - 6.8
3-Nitro-2-pentanol
0.58
3-nitropropionate
-
pH 7.4, 30C
10
butyl-1-nitronate
in 50 mM Tris-HCl, pH 8 at 30C
3.1 - 20
ethyl nitronate
11 - 15.9
ethylnitronate
0.00133
FAD
-
-
1.4
hexyl-1-nitronate
in 50 mM Tris-HCl, pH 8 at 30C
0.9
Nitrocyclohexane
-
-
1 - 29
nitroethane
0.005 - 0.3
O2
0.06
propionate 3-nitronate
-
pH 5.5, 30C
0.27 - 0.34
propionate-3-nitronate
5.5 - 8.3
propyl-1-nitronate
1.61 - 3.1
propyl-2-nitronate
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.6 - 6.2
1-nitrobutane
1.3 - 1.7
1-nitrohexane
3 - 4.3
1-nitropentane
8.3 - 1300
1-Nitropropane
0.82 - 6400
2-Nitropropane
4.4
3-nitropropionate
Neurospora crassa
-
pH 7.4, 30C
42 - 56
butyl-1-nitronate
25 - 4100
ethyl nitronate
16.2 - 185
ethylnitronate
21 - 22
hexyl-1-nitronate
3.5 - 2000
nitroethane
35
pentyl-1-nitronate
Neurospora crassa
Q01284
pH 8.0, 30C
860
propionate 3-nitronate
Cyberlindnera saturnus
-
pH 5.5, 30C
430 - 775
propionate-3-nitronate
42 - 1300
propyl-1-nitronate
15 - 6400
propyl-2-nitronate
additional information
additional information
Neurospora crassa
Q01284
kcat at different conditions, overview, effects of presence of superoxide dimutase or catalase on the turnover rate of the enzyme
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.35
1-nitrobutane
Neurospora crassa
Q01284
in 50 mM Tris-HCl, pH 8 at 30C
6780
1.1
1-nitrohexane
Neurospora crassa
Q01284
in 50 mM Tris-HCl, pH 8 at 30C
4953
0.61
1-nitropentane
Neurospora crassa
Q01284
in 50 mM Tris-HCl, pH 8 at 30C
13377
0.83
1-Nitropropane
Neurospora crassa
Q01284
in 50 mM Tris-HCl, pH 8 at 30C
3300
7.6
3-nitropropionate
Neurospora crassa
-
pH 7.4, 30C
2128
5.6 - 112
butyl-1-nitronate
11537
2.5 - 300
ethyl nitronate
5373
1.02 - 129
ethylnitronate
4420
16 - 130
hexyl-1-nitronate
17654
0.04 - 0.56
nitroethane
1093
0.001 - 11000
O2
9
1.2 - 121
pentyl-1-nitronate
23201
14330
propionate 3-nitronate
Cyberlindnera saturnus
-
pH 5.5, 30C
194797
1300 - 2800
propionate-3-nitronate
150954
11.4 - 170
propyl-1-nitronate
6856
1.6
propyl-2-nitronate
Cyberlindnera mrakii
-
-
5780
additional information
additional information
Neurospora crassa
-
investigation of the pH effects on the kcat/Km-values
2
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.1
3-nitro-1-propionate
-
; competitive inhibitor, pH 7.0, 30C
1.2 - 100
ethylnitronate
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
13
cell free extract
49
after DEAE-Sepharose column chromatography purification only
80
after DEAE-Sepharose column chromatography and octyl-Sepharose column chromatography purification; after DEAE-Sepharose column chromatography and octyl-Sepharose column chromatography purification; at 25C assay temperature, purified recombinant enzyme, substrate ethyl nitronate; at 25C assay temperature, purified recombinant enzyme, substrate ethyl nitronate
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5
-
assay at
7.4
-
assay at
7.4 - 8
-
assay at
7.4 - 8
-
assay at
9.5
-
assay at
additional information
-
when the enzyme is dialyzed against 10 mM potassium phosphate buffer (pH 7.0) immediately after reduction by dithionite, the absorption spectrum similar to that of the native enzyme appears with concomitant restoration of approximately 80% of the activity
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 10
-
assays are carried out at pH 6, 8, 9.5, and 10
6 - 9
-
investigation of the pH effects on the kcat/Km-values
7 - 8.5
-
; when the enzyme is acidified to pH 3.0 and treated in the same way, the prosthetic groups do not dissociate from the protein and almost full activity remained
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 45
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Shewanella oneidensis (strain MR-1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25000
-
1 * 25000 + 1 * 39000, SDS-PAGE
37000
-
dynamic light scattering analysis
39000
-
1 * 25000 + 1 * 39000, SDS-PAGE
39920
calculated; calculated from the amino acid composition of the enzyme
47000
-
4 * 47000, SDS-PAGE
60000
-
gel filtration
64000
-
equilibrium sedimentation
70000
-
gel filtration
80000
recombinant enzyme, gel filtration
185000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
2 * 40000, gel filtration, heat denaturation, and mass spectroscopic analysis
monomer
tetramer
-
4 * 47000, SDS-PAGE
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystallization of the substrate and the complex
-
hanging drop vapour diffusion method using 10% (w/v) polyethylene glycol monomethyl ether 5000
-
crystal structure of NAO from Streptomyces ansochromogenes is determined. It consists of two domains, a TIM barrel domain bound to FMN and C-terminal domain with a novel folding pattern
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 10
-
stable
639225
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40
-
5 min, no loss of activity
45
-
5 min, 14% loss of activity
50
-
5 min, 21.5% loss of activity
55
-
5 min, 60.6% loss of activity
60
-
5 min, 91% loss of activity
65
-
5 min, complete inactivation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
denaturation by sodium dithionite
-
gradual and irreversible loss of activity after treatment with 1% sodium lauryl sulfate or 6 M guanidine HCl, activity is reduced to 50% of its initial activity after 50 min at 37C, complete loss of activity after 10 h
-
immediate denaturation with 8 M urea or thiourea
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 1 mM or 10 mM potassium phosphate buffer, pH 7.0, at least 3 months
-
-20C, 50 mM potassium phosphate, pH 7.4, 6 months, no loss of activity
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; to homogeneity
DEAE fast flow column chromatography and Hi-Prep 16/10 octyl fast flow column chromatography; DEAE fast flow column chromatography and Hi-Prep 16/10 octyl fast flow column chromatography; recombinant enzyme from Escherichia coli strain BL21(DE3) by ammonium sulfate fractionation, DEAE and octyl resin chromatography; recombinant enzyme from Escherichia coli strain BL21(DE3) by ammonium sulfate fractionation, DEAE and octyl resin chromatography
HiLoad XK 16 Superdex 200 prep-grade column gel filtration and Mono Q HR5/5 column chromatography
-
purified to homogeneity
-
to homogeneity
-
using Ni-NTA chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed as a His-tagged fusion protein in Escherichia coli
-
expressed in Escherichia coli
expressed in Escherichia coli BL21(DE3) cells; gene ncd-2, expression in Escherichia coli strain BL21(DE3), subcloning in strain XL-1 Blue; gene ncd-2, expression in Escherichia coli strain BL21(DE3), subcloning in strain XL-1 Blue; expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli C41(DE3) cells
-
expression in Escherichia coli
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
negatively regulated by the transcriptional repressor KstR
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
H199A
-
mutant shows no enzymatic function
H196N
-
site-directed mutagenesis, does to catalyze the formation of ethylnitronate from nitroethane. It is a better catalyst than the wild-type enzyme for oxidative turnover with ethylnitronate; the H196N variant form of the enzyme does to catalyze the formation of ethylnitronate from nitroethane. The H196N variant is a better catalyst than the wild-type enzyme for oxidative turnover with ethylnitronate
H152A
-
His152 likely functions as the catalytic base that initiates oxidation of neutral substrates by abstracting a proton from the alpha-carbon; site-directed mutagenesis. His152 likely functions as the catalytic base that initiates oxidation of neutral substrates by abstracting a proton from the alpha-carbon
S288A
-
mutant enzyme forms inclusion bodies when overexpressed in Escherichia coli; site-directed mutagenesis. Mutant enzyme forms inclusion bodies when overexpressed in Escherichia coli
H179D
mutant enzyme shows no activity. Crystal structure of mutant H179D is determined: The structural superimposition of wild-type Sa-NAO and mutant H179D-nitroethane shows no significant structural variation
H179K
mutant enzyme shows no activity
H179V
mutant enzyme shows no activity
Show AA Sequence (1567 entries)
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