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Literature summary for 1.13.12.16 extracted from

  • Salvi, F.; Agniswamy, J.; Yuan, H.; Vercammen, K.; Pelicaen, R.; Cornelis, P.; Spain, J.C.; Weber, I.T.; Gadda, G.
    The combined structural and kinetic characterization of a bacterial nitronate monooxygenase from Pseudomonas aeruginosa PAO1 establishes NMO class I and II (2014), J. Biol. Chem., 289, 23764-23775 .
    View publication on PubMedView publication on EuropePMC
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Cloned(Commentary)

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 (Commentary)

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 [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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Pseudomonas aeruginosa Q9HWH9
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain Rosetta(DE3)pLysS by affinity chromatography Pseudomonas aeruginosa

Reaction

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
a

Substrates and Products (Substrate)

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

Subunits Comment Organism
additional information 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

Synonyms Comment Organism
NMO
-
 Pseudomonas aeruginosa
Pa-NMO
-
 Pseudomonas aeruginosa
PA4202
-
 Pseudomonas aeruginosa
protein PA4202
-
 Pseudomonas aeruginosa

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Pseudomonas aeruginosa

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Pseudomonas aeruginosa

Cofactor

Cofactor Comment Organism Structure
FMN dependent on, FMN-binding domain structure, overview Pseudomonas aeruginosa

General Information

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 [mM/s]

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