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Literature summary for 1.7.3.1 extracted from

  • Fitzpatrick, P.F.; Bozinovski, D.M.; Heroux, A.; Shaw, P.G.; Valley, M.P.; Orville, A.M.
    Mechanistic and structural analyses of the roles of Arg409 and Asp402 in the reaction of the flavoprotein nitroalkane oxidase (2007), Biochemistry, 46, 13800-13808.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.7.3.1

Crystallization (Commentary)

Crystallization (Comment) Organism
hexagonal rod-shaped crystals of R409K and D402E NAO were obtained using hanging drop vapor-diffusion methods Fusarium oxysporum

Protein Variants

Protein Variants Comment Organism
D402E mutation results in a decrease in the rate constant for proton abstraction of 18fold. The structure of the D402E enzyme, determined at 2.4 A resolution, shows that there is a smaller increase in the distance between Arg409 and the carboxylate at position 402. The interaction of this residue with Ser276 is perturbed Fusarium oxysporum
D402E the mutation results in a decrease in the rate constant for proton abstraction of 18fold. The structure of the D402E enzyme shows that there is a smaller increase in the distance between Arg409 and the carboxylate at position 402 (compared to mutant enzyme R409K), and the interaction of this residue with Ser276 is perturbed Fusarium oxysporum
R409K the mutation results in a decrease in the rate constant for proton abstraction of 100fold. Analysis of the three-dimensional structure of the R409K enzyme shows that the critical structural change is an increase in the distance between the carboxylate of Asp402 and the positively charged nitrogen in the side chain of the residue at position 409 Fusarium oxysporum
R409K the mutation results in a decrease in the rate constant for proton abstraction of 100fold. Analysis of the three-dimensional structure of the R409K enzyme, determined by X-ray crystallography to a resolution of 2.65 A, shows that the critical structural change is an increase in the distance between the carboxylate of Asp402 and the positively charged nitrogen in the side chain of the residue at position 409 Fusarium oxysporum

Inhibitors

Inhibitors Comment Organism Structure
nitroethane substrate inhibition detected in mutant enzyme R409K, no substrate inhibition is detected in mutant enzyme D402E Fusarium oxysporum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.019
-
 O2 pH 8.0, 30°C, mutant enzyme D402E Fusarium oxysporum
0.042
-
 O2 pH 8.0, 30°C, mutant enzyme R409K Fusarium oxysporum
0.042
-
 nitroethane pH 8.0, 30°C, mutant enzyme R409K Fusarium oxysporum
26.3
-
 nitroethane pH 8.0, 30°C, mutant enzyme R409K Fusarium oxysporum

Organism

Organism UniProt Comment Textmining
Fusarium oxysporum
-
-
-
Fusarium oxysporum
-
 recombinant
-

Reaction

Reaction Comment Organism Reaction ID
ethylnitronate + O2 + FMNH2 = acetaldehyde + nitrite + FMN + H2O critical importance of the interaction between Asp402 and Arg409 for proton abstraction by nitroalkane oxidase Fusarium oxysporum
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
nitroethane + H2O + O2
-
 Fusarium oxysporum acetaldehyde + nitrite + H2O2
-
 ?
nitroethane + H2O + O2
-
 Fusarium oxysporum ?
-
 ?

Synonyms

Synonyms Comment Organism
NAO
-
 Fusarium oxysporum
nitroalkane oxidase
-
 Fusarium oxysporum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.6
-
 O2 pH 8.0, 30°C, mutant enzyme R409K Fusarium oxysporum
2.6
-
 nitroethane pH 8.0, 30°C, mutant enzyme R409K Fusarium oxysporum

Cofactor

Cofactor Comment Organism Structure
FAD
-
 Fusarium oxysporum

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
152
-
 nitroethane pH 8.0, 30°C, mutant enzyme R409K, substrate inhibition constant Fusarium oxysporum