Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes | Fusarium oxysporum |
Protein Variants | Comment | Organism |
---|---|---|
D402A | site-directed mutation of the active site base, 20fold reduced catalytic efficiency with neutral nitroethane as substrate compared to the wild-type enzyme, while the wild-type enzyme prefers the neutral substrate the mutant prefers the anion substrate form, altered pH-dependence with both substrate forms compared to the wild-type enzyme | Fusarium oxysporum |
D402N | site-directed mutation of the active site base, 140fold reduced catalytic efficiency with neutral nitroethane as substrate compared to the wild-type enzyme, while the wild-type enzyme prefers the neutral substrate the mutant prefers the anion substrate form, altered pH-dependence with both substrate forms compared to the wild-type enzyme | Fusarium oxysporum |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Fusarium oxysporum | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes | Fusarium oxysporum |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ethylnitronate + O2 + FMNH2 = acetaldehyde + nitrite + FMN + H2O | Asp402 acts as the active site base of the enzyme, mechanism: catalysis is initiated by the active site base Asp402 that deprotonates the neutral substrate to yield a carbanion that can attack the N-5 position of the oxidized flavin, CH-bond cleavage is rate limiting for the reduction of the enzyme by nitroethane | Fusarium oxysporum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | enzyme catalyzes the oxidation of nitroalkanes to the respective aldehydes or ketones with consumption of molecular oxygen and releasing nitrite and hydrogen peroxide | Fusarium oxysporum | ? | - |
? | |
nitroethane + O2 + H2O | the neutral substrate form is preferred, 5fold lower activity with the substrate anion, the D402 mutant enzymes in contrast prefer the anionic substrate form | Fusarium oxysporum | ethanal + nitrite + H2O2 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
NAO | - |
Fusarium oxysporum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Fusarium oxysporum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
the pH-dependence of the reaction with the wild-type enzyme depends on the substrate form, at pH 7.4 activity with the neutral substrate increases while is decreases with the nitroethane anion | Fusarium oxysporum |
8 | - |
assay at | Fusarium oxysporum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Fusarium oxysporum |