Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Podospora anserina |
Crystallization (Comment) | Organism |
---|---|
vapor diffusion method, using 2.5 M magnesium sulfate, 0.1 M 2-(N-morpholino) ethanesulfonic acid buffer, pH 6.0, and 18% (v/v) glycerol | Podospora anserina |
Protein Variants | Comment | Organism |
---|---|---|
D399N | the mutation decreases the kcat/KM value for nitroethane over 2 orders of magnitude | Podospora anserina |
R406K | the mutation decreases the kcat/KM value for nitroethane about 64fold | Podospora anserina |
S373A | the mutation decreases the kcat/KM value for nitroethane about 3fold | Podospora anserina |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.39 | - |
O2 | wild type enzyme, in 50 mM HEPES, pH 8.0, 30°C | Podospora anserina | |
10 | - |
nitroethane | mutant enzyme D399N, in 50 mM HEPES, pH 8.0, 30°C | Podospora anserina | |
13 | - |
nitroethane | wild type enzyme, in 50 mM HEPES, pH 8.0, 30°C | Podospora anserina | |
24 | - |
nitroethane | mutant enzyme S273A, in 50 mM HEPES, pH 8.0, 30°C | Podospora anserina | |
25 | - |
nitroethane | mutant enzyme R406K, in 50 mM HEPES, pH 8.0, 30°C | Podospora anserina |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Podospora anserina | B2AM55 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography | Podospora anserina |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
nitroethane + O2 + H2O | - |
Podospora anserina | ethanal + nitrite + H2O2 + H+ | - |
ir |
Synonyms | Comment | Organism |
---|---|---|
NAO | - |
Podospora anserina |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.037 | - |
nitroethane | mutant enzyme D399N, in 50 mM HEPES, pH 8.0, 30°C | Podospora anserina | |
5 | - |
nitroethane | mutant enzyme S273A, in 50 mM HEPES, pH 8.0, 30°C | Podospora anserina | |
9 | - |
nitroethane | mutant enzyme R406K, in 50 mM HEPES, pH 8.0, 30°C | Podospora anserina | |
15 | - |
O2 | wild type enzyme, in 50 mM HEPES, pH 8.0, 30°C | Podospora anserina | |
15 | - |
nitroethane | wild type enzyme, in 50 mM HEPES, pH 8.0, 30°C | Podospora anserina |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | 0.5 mM FAD is used in assay conditions | Podospora anserina |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.004 | - |
nitroethane | mutant enzyme D399N, in 50 mM HEPES, pH 8.0, 30°C | Podospora anserina | |
0.21 | - |
nitroethane | mutant enzyme S273A, in 50 mM HEPES, pH 8.0, 30°C | Podospora anserina | |
0.37 | - |
nitroethane | mutant enzyme R406K, in 50 mM HEPES, pH 8.0, 30°C | Podospora anserina | |
0.74 | - |
nitroethane | wild type enzyme, in 50 mM HEPES, pH 8.0, 30°C | Podospora anserina | |
38 | - |
O2 | wild type enzyme, in 50 mM HEPES, pH 8.0, 30°C | Podospora anserina |