Cloned (Comment) | Organism |
---|---|
gene pa4202, DNA and amino acid sequence determination and analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain Rosetta(DE3)pLysS | Pseudomonas aeruginosa |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His-tagged enzyme, vapor diffusion hanging drop method at room temperature, 0.002 ml of 14 mg/ml protein in 50 mM Tris-Cl, pH 8.0, 100 mM NaCl, 20 mM 2-nitropropane, with 0.002 ml of reservoir solution containing 14% w/v PEG 5000 monomethylether and 0.1 M Na-HEPES, pH 7.0, and equilibration against 1 ml of reservoir solution, 7-10 days, X-ray diffraction structure determination and analysis at 1.44 A resolution, molecular replacement | Pseudomonas aeruginosa |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetic mechanism | Pseudomonas aeruginosa | |
0.11 | - |
propionate-3-nitronate | pH 7.5, 30°C, recombinant enzyme | Pseudomonas aeruginosa | |
5 | - |
ethylnitronate | pH 7.5, 30°C, recombinant enzyme | Pseudomonas aeruginosa | |
6 | - |
propyl-1-nitronate | pH 7.5, 30°C, recombinant enzyme | Pseudomonas aeruginosa |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
3 propionate-3-nitronate + O2 | Pseudomonas aeruginosa | - |
3 malonic semialdehyde + nitrite + 2 nitrate + H2O2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas aeruginosa | Q9HWH9 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain Rosetta(DE3)pLysS by affinity chromatography | Pseudomonas aeruginosa |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ethylnitronate + O2 = acetaldehyde + nitrite + other products | catalyze the oxidation of propionate-3-nitronate or other nitronate analogues through a radical mechanism involving a catalytic flavosemiquinone without formation of a canonical C4a-(hydro)peroxyflavin | Pseudomonas aeruginosa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3 propionate-3-nitronate + O2 | - |
Pseudomonas aeruginosa | 3 malonic semialdehyde + nitrite + 2 nitrate + H2O2 | - |
? | |
3 propionate-3-nitronate + O2 | best substrate | Pseudomonas aeruginosa | 3 malonic semialdehyde + nitrite + 2 nitrate + H2O2 | - |
? | |
butyl-1-nitronate + O2 | - |
Pseudomonas aeruginosa | ? | - |
? | |
ethylnitronate + O2 | - |
Pseudomonas aeruginosa | acetaldehyde + nitrite + other products | - |
? | |
additional information | the enzyme is active on propionate-3-nitronate and other alkyl nitronates, but cannot oxidize nitroalkanes, e.g. 3-nitropropionate, nitroethane, 1-nitropropane, 1-nitrobutane, 1-nitropentane, or 2-nitropropane. Anaerobic reduction of the enzyme with propionate-3-nitronate yields a flavosemiquinone | Pseudomonas aeruginosa | ? | - |
? | |
pentyl-1-nitronate + O2 | - |
Pseudomonas aeruginosa | ? | - |
? | |
propyl-1-nitronate + O2 | - |
Pseudomonas aeruginosa | ? | - |
? | |
propyl-2-nitronate + O2 | - |
Pseudomonas aeruginosa | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the dimer interface includes eight contacts mainly in the FMN-binding domain and it does not seem to be directly relevant for catalysis as it is far from the active site pocket, structure analysis, overview | Pseudomonas aeruginosa |
Synonyms | Comment | Organism |
---|---|---|
NMO | - |
Pseudomonas aeruginosa |
Pa-NMO | - |
Pseudomonas aeruginosa |
PA4202 | - |
Pseudomonas aeruginosa |
protein PA4202 | - |
Pseudomonas aeruginosa |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Pseudomonas aeruginosa |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
350 | - |
ethylnitronate | pH 7.5, 30°C, recombinant enzyme | Pseudomonas aeruginosa | |
1120 | - |
propyl-1-nitronate | pH 7.5, 30°C, recombinant enzyme | Pseudomonas aeruginosa | |
1300 | - |
propionate-3-nitronate | pH 7.5, 30°C, recombinant enzyme | Pseudomonas aeruginosa |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Pseudomonas aeruginosa |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FMN | dependent on, FMN-binding domain structure, overview | Pseudomonas aeruginosa |
General Information | Comment | Organism |
---|---|---|
evolution | NMO is a member of the group H flavin-dependent monooxygenases | Pseudomonas aeruginosa |
additional information | four conserved motifs are identified in PA4202. Oxidation of propionate-3-nitronate by NMO yields malonic semialdehyde, which is an important metabolite that can be converted to acetyl-CoA, acetate, or 3-hydroxypropionate and enter various catabolic or anabolic pathways. Active site structure analysis, overview | Pseudomonas aeruginosa |
physiological function | nitronate monooxygenase oxidizes the mitochondrial toxin propionate 3-nitronate to malonate semialdehyde, and detoxifies the deadly toxin | Pseudomonas aeruginosa |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
29 | - |
propyl-2-nitronate | pH 7.5, 30°C, recombinant enzyme | Pseudomonas aeruginosa | |
33 | - |
butyl-1-nitronate | pH 7.5, 30°C, recombinant enzyme | Pseudomonas aeruginosa | |
70 | - |
ethylnitronate | pH 7.5, 30°C, recombinant enzyme | Pseudomonas aeruginosa | |
91 | - |
pentyl-1-nitronate | pH 7.5, 30°C, recombinant enzyme | Pseudomonas aeruginosa | |
186.71 | - |
propyl-1-nitronate | pH 7.5, 30°C, recombinant enzyme | Pseudomonas aeruginosa | |
11818.2 | - |
propionate-3-nitronate | pH 7.5, 30°C, recombinant enzyme | Pseudomonas aeruginosa |