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Literature summary for 1.7.1.17 extracted from

  • Cao, X.; Di, M.; Wang, J.
    Expansion of the active site of the azoreductase from Shewanella oneidensis MR-1 (2017), J. Mol. Graph. Model., 78, 213-220 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Shewanella oneidensis

Protein Variants

Protein Variants Comment Organism
F117A the mutant shows 208% activity with Methyl Red and 176% activity with NADH compared to the wild type enzyme Shewanella oneidensis
F161A inactive Shewanella oneidensis
G140F inactive Shewanella oneidensis
G141F inactive Shewanella oneidensis
H143A the mutant shows 19% activity with Methyl Red and 76% activity with NADH compared to the wild type enzyme Shewanella oneidensis
N96A the mutant shows 9% activity with Methyl Red and 37% activity with NADH compared to the wild type enzyme Shewanella oneidensis
R139A the mutant shows 63% activity with Methyl Red and 78% activity with NADH compared to the wild type enzyme Shewanella oneidensis
R59A the mutant shows 70% activity with Methyl Red and 83% activity with NADH compared to the wild type enzyme Shewanella oneidensis
S16A the mutant shows 29% activity with Methyl Red and 49% activity with NADH compared to the wild type enzyme Shewanella oneidensis
Y119A the mutant shows 59% activity with Methyl Red and 44% activity with NADH compared to the wild type enzyme Shewanella oneidensis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.01155
-
Methyl red wild type enzyme, at pH 7.4 and 30°C Shewanella oneidensis
0.01603
-
Methyl red mutant enzyme F117A, at pH 7.4 and 30°C Shewanella oneidensis
0.01928
-
Methyl red mutant enzyme Y119A, at pH 7.4 and 30°C Shewanella oneidensis
0.02714
-
NADH mutant enzyme F117A, at pH 7.4 and 30°C Shewanella oneidensis
0.0296
-
Methyl red mutant enzyme R59A, at pH 7.4 and 30°C Shewanella oneidensis
0.04338
-
NADH mutant enzyme H143A, at pH 7.4 and 30°C Shewanella oneidensis
0.04809
-
Methyl red mutant enzyme R139A, at pH 7.4 and 30°C Shewanella oneidensis
0.06872
-
Methyl red mutant enzyme H143A, at pH 7.4 and 30°C Shewanella oneidensis
0.07676
-
Methyl red mutant enzyme S16A, at pH 7.4 and 30°C Shewanella oneidensis
0.09315
-
Methyl red mutant enzyme N96A, at pH 7.4 and 30°C Shewanella oneidensis
0.18777
-
NADH wild type enzyme, at pH 7.4 and 30°C Shewanella oneidensis
0.2241
-
NADH mutant enzyme R139A, at pH 7.4 and 30°C Shewanella oneidensis
0.3158
-
NADH mutant enzyme N96A, at pH 7.4 and 30°C Shewanella oneidensis
0.3433
-
NADH mutant enzyme R59A, at pH 7.4 and 30°C Shewanella oneidensis
0.5307
-
NADH mutant enzyme S16A, at pH 7.4 and 30°C Shewanella oneidensis
0.6551
-
NADH mutant enzyme Y119A, at pH 7.4 and 30°C Shewanella oneidensis

Organism

Organism UniProt Comment Textmining
Shewanella oneidensis Q8E990
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Shewanella oneidensis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Methyl Red + NADH + H+
-
Shewanella oneidensis 2-aminobenzoic acid + N,N-dimethyl-1,4-phenylenediamine + NAD+
-
?

Synonyms

Synonyms Comment Organism
AzoR
-
Shewanella oneidensis
azoreductase
-
Shewanella oneidensis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2900
-
Methyl red mutant enzyme N96A, at pH 7.4 and 30°C Shewanella oneidensis
6350
-
Methyl red mutant enzyme H143A, at pH 7.4 and 30°C Shewanella oneidensis
9800
-
Methyl red mutant enzyme S16A, at pH 7.4 and 30°C Shewanella oneidensis
14420
-
NADH mutant enzyme N96A, at pH 7.4 and 30°C Shewanella oneidensis
17170
-
NADH mutant enzyme Y119A, at pH 7.4 and 30°C Shewanella oneidensis
19180
-
NADH mutant enzyme S16A, at pH 7.4 and 30°C Shewanella oneidensis
20020
-
Methyl red mutant enzyme Y119A, at pH 7.4 and 30°C Shewanella oneidensis
21480
-
Methyl red mutant enzyme R139A, at pH 7.4 and 30°C Shewanella oneidensis
23680
-
Methyl red mutant enzyme R59A, at pH 7.4 and 30°C Shewanella oneidensis
29390
-
NADH mutant enzyme H143A, at pH 7.4 and 30°C Shewanella oneidensis
30200
-
NADH mutant enzyme R139A, at pH 7.4 and 30°C Shewanella oneidensis
32130
-
NADH mutant enzyme R59A, at pH 7.4 and 30°C Shewanella oneidensis
33860
-
Methyl red wild type enzyme, at pH 7.4 and 30°C Shewanella oneidensis
33900
-
NADH wild type enzyme, at pH 7.4 and 30°C Shewanella oneidensis
68230
-
NADH mutant enzyme F117A, at pH 7.4 and 30°C Shewanella oneidensis
70360
-
Methyl red mutant enzyme F117A, at pH 7.4 and 30°C Shewanella oneidensis

Cofactor

Cofactor Comment Organism Structure
FMN
-
Shewanella oneidensis
NADH
-
Shewanella oneidensis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.03
-
Methyl red mutant enzyme N96A, at pH 7.4 and 30°C Shewanella oneidensis
0.03
-
NADH mutant enzyme Y119A, at pH 7.4 and 30°C Shewanella oneidensis
0.04
-
NADH mutant enzyme S16A, at pH 7.4 and 30°C Shewanella oneidensis
0.05
-
NADH mutant enzyme N96A, at pH 7.4 and 30°C Shewanella oneidensis
0.07
-
NADH mutant enzyme H143A, at pH 7.4 and 30°C Shewanella oneidensis
0.09
-
Methyl red mutant enzyme H143A, at pH 7.4 and 30°C Shewanella oneidensis
0.09
-
NADH mutant enzyme R59A, at pH 7.4 and 30°C Shewanella oneidensis
0.13
-
Methyl red mutant enzyme S16A, at pH 7.4 and 30°C Shewanella oneidensis
0.14
-
NADH mutant enzyme R139A, at pH 7.4 and 30°C Shewanella oneidensis
0.18
-
NADH wild type enzyme, at pH 7.4 and 30°C Shewanella oneidensis
0.45
-
Methyl red mutant enzyme R139A, at pH 7.4 and 30°C Shewanella oneidensis
0.8
-
Methyl red mutant enzyme R59A, at pH 7.4 and 30°C Shewanella oneidensis
1.04
-
Methyl red mutant enzyme Y119A, at pH 7.4 and 30°C Shewanella oneidensis
2.51
-
NADH mutant enzyme F117A, at pH 7.4 and 30°C Shewanella oneidensis
2.93
-
Methyl red wild type enzyme, at pH 7.4 and 30°C Shewanella oneidensis
4.39
-
Methyl red mutant enzyme F117A, at pH 7.4 and 30°C Shewanella oneidensis