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Literature summary for 1.13.12.16 extracted from

  • Francis, K.; Gadda, G.
    The nonoxidative conversion of nitroethane to ethylnitronate in Neurospora crassa 2-nitropropane dioxygenase is catalyzed by Histidine 196 (2008), Biochemistry, 47, 9136-9144.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Neurospora crassa

Protein Variants

Protein Variants Comment Organism
H196N 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 Neurospora crassa
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 Neurospora crassa

Inhibitors

Inhibitors Comment Organism Structure
ethylnitronate
-
Neurospora crassa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information Km-value for O2 (wild-type enzyme) is below 0.005 mM Neurospora crassa
0.005
-
O2 30°C, pH 8, wild-type enzyme, ethylnitronate as a substrate Neurospora crassa
0.034
-
O2 30°C, pH 8, mutant enzyme H196N Neurospora crassa
0.034
-
O2 30°C, pH 8, mutant enzyme H196N, ethylnitronate as a substrate Neurospora crassa
11
-
ethylnitronate 30°C, pH 9.5, mutant enzyme H196N Neurospora crassa
15.9
-
ethylnitronate 30°C, pH 9.5, wild-type enzyme Neurospora crassa

Organism

Organism UniProt Comment Textmining
Neurospora crassa
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Neurospora crassa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ethyl nitronate + O2
-
Neurospora crassa ? + nitrite
-
?
additional information 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 Neurospora crassa ?
-
?
nitroethane + O2 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 Neurospora crassa acetaldehyde + nitrite
-
?
nitroethane + O2 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 Neurospora crassa ethylnitronate
-
r
nitroethane + O2
-
Neurospora crassa ? + nitrite
-
?

Synonyms

Synonyms Comment Organism
2-nitropropane dioxygenase
-
Neurospora crassa

Temperature Optimum [°C]

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

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
16.2
-
ethylnitronate 30°C, pH 9.5, wild-type enzyme Neurospora crassa
185
-
ethylnitronate 30°C, pH 9.5, mutant enzyme H196N Neurospora crassa

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9.5
-
assay at Neurospora crassa

pH Range

pH Minimum pH Maximum Comment Organism
6 10 assays are carried out at pH 6, 8, 9.5, and 10 Neurospora crassa

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.2
-
ethylnitronate 30°C, pH 9.5, mutant enzyme H196N Neurospora crassa
100
-
ethylnitronate 30°C, pH 9.5, wild-type enzyme Neurospora crassa

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.001
-
O2 30°C, pH 8, wild-type enzyme Neurospora crassa
0.0055
-
O2 30°C, pH 8, mutant enzyme H196N Neurospora crassa
1.02
-
ethylnitronate 30°C, pH 8, wild-type enzyme Neurospora crassa
16.8
-
ethylnitronate 30°C, pH 8, mutant enzyme H196N Neurospora crassa