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Literature summary for 1.13.12.16 extracted from

  • Francis, K.; Russell, B.; Gadda, G.
    Involvement of a flavosemiquinone in the enzymatic oxidation of nitroalkanes catalyzes by 2-nitropropane dioxygenase (2005), J. Biol. Chem., 280, 5195-5204.
    View publication on PubMed
Search for more literature for 1.13.12.16

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Neurospora crassa
gene ncd-2, expression in Escherichia coli strain BL21(DE3), subcloning in strain XL-1 Blue Neurospora crassa
gene ncd-2, expression in Escherichia coli strain BL21(DE3), subcloning in strain XL-1 Blue; expressed in Escherichia coli BL21(DE3) cells Neurospora crassa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information kinetics, steady-state kinetic mechanism Neurospora crassa
1.4
-
 1-nitrohexane in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
1.4
-
 hexyl-1-nitronate in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
3.1
-
 ethyl nitronate in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
5.5
-
 propyl-1-nitronate in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
7.1
-
 1-nitropentane in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
10
-
 butyl-1-nitronate in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
15
-
 1-nitrobutane in 50 mM Tris-Cl, pH 8 at 30°C Neurospora crassa
17.9
-
 1-nitrobutane in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
24
-
 1-Nitropropane in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
29
-
 nitroethane in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa

Metals/Ions

Metals/Ions Comment Organism Structure
Iron stoichiometry of 0.02 FMN per 1 monomer of enzyme Neurospora crassa
additional information devoid of iron Neurospora crassa

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
39920
-
 calculated Neurospora crassa
39920
-
 calculated from the amino acid composition of the enzyme Neurospora crassa
40000
-
 SDS-PAGE Neurospora crassa
80000
-
 recombinant enzyme, gel filtration Neurospora crassa

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Neurospora crassa enzyme is more specific for nitronates than nitroalkanes ?
-
 ?

Organism

Organism UniProt Comment Textmining
Neurospora crassa Q01284
-
-
Neurospora crassa Q01284 gene ncd-2
-

Purification (Commentary)

Purification (Comment) Organism
DEAE fast flow column chromatography and Hi-Prep 16/10 octyl fast flow column chromatography Neurospora crassa
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 Neurospora crassa
recombinant enzyme from Escherichia coli strain BL21(DE3) by ammonium sulfate fractionation, DEAE and octyl resin chromatography Neurospora crassa

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
13
-
 cell free extract Neurospora crassa
49
-
 after DEAE-Sepharose column chromatography purification only Neurospora crassa
80
-
 at 25°C assay temperature, purified recombinant enzyme, substrate ethyl nitronate Neurospora crassa
80
-
 after DEAE-Sepharose column chromatography and octyl-Sepharose column chromatography purification Neurospora crassa
80
-
 after DEAE-Sepharose column chromatography and octyl-Sepharose column chromatography purification; at 25°C assay temperature, purified recombinant enzyme, substrate ethyl nitronate Neurospora crassa

Storage Stability

Storage Stability Organism
-20°C, 50 mM potassium phosphate, pH 7.4, 6 months, no loss of activity Neurospora crassa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1-nitrobutane + O2
-
 Neurospora crassa butyraldehyde + HNO2
-
 ?
1-nitrobutane + O2 neutral and anionic form Neurospora crassa butyraldehyde + HNO2
-
 ?
1-nitrobutane + O2
-
 Neurospora crassa butyraldehyde + nitrite
-
 ?
1-nitrohexane + O2
-
 Neurospora crassa hexanaldehyde + HNO2
-
 ?
1-nitrohexane + O2 neutral and anionic form Neurospora crassa hexanaldehyde + HNO2
-
 ?
1-nitrohexane + O2
-
 Neurospora crassa hexanaldehyde + nitrite
-
 ?
1-nitropentane + O2
-
 Neurospora crassa pentanaldehyde + HNO2
-
 ?
1-nitropentane + O2 neutral and anionic form Neurospora crassa pentanaldehyde + HNO2
-
 ?
1-nitropentane + O2
-
 Neurospora crassa pentanaldehyde + nitrite
-
 ?
1-nitropropane + O2
-
 Neurospora crassa propionaldehyde + HNO2
-
 ?
1-nitropropane + O2 neutral and anionic form Neurospora crassa propionaldehyde + HNO2
-
 ?
1-nitropropane + O2
-
 Neurospora crassa propionaldehyde + nitrite
-
 ?
2-nitropropane + O2
-
 Neurospora crassa acetone + HNO2
-
 ?
2-nitropropane + O2 neutral and anionic form Neurospora crassa acetone + HNO2
-
 ?
2-nitropropane + O2
-
 Neurospora crassa acetone + nitrite
-
 ?
butyl-1-nitronate + O2
-
 Neurospora crassa ? + nitrite
-
 ?
butyl-1-nitronate + O2 neutral and anionic form Neurospora crassa ? + nitrite
-
 ?
ethyl nitronate + O2
-
 Neurospora crassa ? + nitrite
-
 ?
ethylnitronate + O2 neutral and anionic form Neurospora crassa acetaldehyde + HNO2
-
 ?
hexyl-1-nitronate + O2
-
 Neurospora crassa ? + nitrite
-
 ?
hexyl-1-nitronate + O2 neutral and anionic form Neurospora crassa ? + nitrite
-
 ?
additional information the reduced enzyme can reduce the substrate under anaerobically conditions, substrate specificity with nitroalkanes and alkyl nitronates, O2 is delivered from air-saturated buffer in the assay reaction, enzyme catalyzes the 2-step oxidative denitrification of nitroalkanes to their corresponding carbonyl compounds and nitrite Neurospora crassa ?
-
 ?
additional information enzyme is more specific for nitronates than nitroalkanes Neurospora crassa ?
-
 ?
nitroethane + O2
-
 Neurospora crassa ethanal + nitrite
-
 ?
nitroethane + O2 neutral and anionic form Neurospora crassa ethanal + nitrite
-
 ?
nitroethane + O2 + H2O
-
 Neurospora crassa ethanal + nitrite + H2O2
-
 ?
pentyl-1-nitronate + O2
-
 Neurospora crassa ? + nitrite
-
 ?
pentyl-1-nitronate + O2 neutral and anionic form Neurospora crassa ? + nitrite
-
 ?
propyl-1-nitronate + O2
-
 Neurospora crassa ? + nitrite
-
 ?
propyl-1-nitronate + O2 neutral and anionic form Neurospora crassa ? + nitrite
-
 ?
propyl-1-nitronate + O2
-
 Neurospora crassa ?
-
 ?
propyl-2-nitronate + O2
-
 Neurospora crassa ? + nitrite
-
 ?
propyl-2-nitronate + O2 neutral and anionic form Neurospora crassa ? + nitrite
-
 ?

Subunits

Subunits Comment Organism
dimer 2 * 40000, recombinant enzyme, SDS-PAGE Neurospora crassa
homodimer 2 * 40000, gel filtration, heat denaturation, and mass spectroscopic analysis Neurospora crassa

Synonyms

Synonyms Comment Organism
2-nitropropane dioxygenase
-
 Neurospora crassa

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
-
 Neurospora crassa
30
-
 assay at Neurospora crassa

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
additional information
-
 additional information kcat at different conditions, overview, effects of presence of superoxide dimutase or catalase on the turnover rate of the enzyme Neurospora crassa
0.82
-
 2-Nitropropane pH 8.0, 30°C Neurospora crassa
1.3
-
 1-nitrohexane pH 8.0, 30°C Neurospora crassa
1.5
-
 1-nitrohexane in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
1.7
-
 1-nitrohexane pH 8.0, 30°C Neurospora crassa
3
-
 1-nitropentane pH 8.0, 30°C Neurospora crassa
3.5
-
 nitroethane pH 8.0, 30°C Neurospora crassa
3.6
-
 1-nitrobutane pH 8.0, 30°C Neurospora crassa
4.3
-
 1-nitropentane in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
5.9
-
 1-nitrobutane pH 8.0, 30°C Neurospora crassa
6.2
-
 1-nitrobutane in 50 mM Tris-Cl, pH 8 at 30°C Neurospora crassa
8.3
-
 1-Nitropropane pH 8.0, 30°C Neurospora crassa
11
-
 nitroethane pH 8.0, 30°C Neurospora crassa
14
-
 nitroethane in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
15
-
 propyl-2-nitronate pH 8.0, 30°C Neurospora crassa
20
-
 1-Nitropropane in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
21
-
 hexyl-1-nitronate pH 8.0, 30°C Neurospora crassa
22
-
 hexyl-1-nitronate in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
29
-
 propyl-2-nitronate pH 8.0, 30°C Neurospora crassa
35
-
 pentyl-1-nitronate pH 8.0, 30°C Neurospora crassa
40
-
 ethyl nitronate pH 8.0, 30°C Neurospora crassa
42
-
 propyl-1-nitronate pH 8.0, 30°C Neurospora crassa
42
-
 butyl-1-nitronate pH 8.0, 30°C Neurospora crassa
51
-
 ethyl nitronate in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
55
-
 butyl-1-nitronate in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
56
-
 butyl-1-nitronate pH 8.0, 30°C Neurospora crassa
57
-
 ethylnitronate pH 8.0, 30°C Neurospora crassa
60
-
 propyl-1-nitronate in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
-
 Neurospora crassa
8
-
 assay at Neurospora crassa

pH Range

pH Minimum pH Maximum Comment Organism
6 9.5
-
 Neurospora crassa

Cofactor

Cofactor Comment Organism Structure
FMN dependent on, stoichiometry of 0.84 FMN per 1 monomer of enzyme, noncovalently bound Neurospora crassa
FMN contains a mol of non-covalently bound FMN per mol of subunit, contains 0.84 FMN per monomer of enzyme Neurospora crassa
FMN dependent on, stoichiometry of 0.84 FMN per 1 monomer of enzyme, noncovalently bound, contains a mol of non-covalently bound FMN per mol of subunit, contains 0.84 FMN per monomer of enzyme Neurospora crassa

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.35
-
 1-nitrobutane in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
0.48
-
 nitroethane in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
0.61
-
 1-nitropentane in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
0.83
-
 1-Nitropropane in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
1.1
-
 1-nitrohexane in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
1.2
-
 pentyl-1-nitronate in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
5.6
-
 butyl-1-nitronate in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
11.4
-
 propyl-1-nitronate in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
16
-
 hexyl-1-nitronate in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa
16.5
-
 ethyl nitronate in 50 mM Tris-HCl, pH 8 at 30°C Neurospora crassa