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Literature summary for 1.7.3.1 extracted from

  • Tormos, J.R.; Taylor, A.B.; Daubner, S.C.; Hart, P.J.; Fitzpatrick, P.F.
    Identification of a hypothetical protein from Podospora anserina as a nitroalkane oxidase (2010), Biochemistry, 49, 5035-5041.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Podospora anserina

Crystallization (Commentary)

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

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 [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

Organism UniProt Comment Textmining
Podospora anserina B2AM55
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography Podospora anserina

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
nitroethane + O2 + H2O
-
Podospora anserina ethanal + nitrite + H2O2 + H+
-
ir

Synonyms

Synonyms Comment Organism
NAO
-
Podospora anserina

Turnover Number [1/s]

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

Cofactor Comment Organism Structure
FAD 0.5 mM FAD is used in assay conditions Podospora anserina

kcat/KM [mM/s]

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