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Literature summary for 1.7.1.17 extracted from

  • Romero, E.; Savino, S.; Fraaije, M.W.; Loncar, N.
    Mechanistic and crystallographic studies of azoreductase AzoA from Bacillus wakoensis A01 (2020), ACS Chem. Biol., 15, 504-512 .
    View publication on PubMedView publication on EuropePMC

Crystallization (Commentary)

Crystallization (Comment) Organism
in complex with FMN, sitting drop vapor diffusion method, using 0.17 M ammonium sulfate and 25.2% (w/v) PEG 4000 Alkalihalobacillus wakoensis

Protein Variants

Protein Variants Comment Organism
W60D the mutant retains more than 10% of wild type activity Alkalihalobacillus wakoensis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.025
-
menadione mutant enzyme W60A, at pH 8.5 and 25°C Alkalihalobacillus wakoensis
0.061
-
1,2-naphthoquinone wild type enzyme, at pH 8.5 and 25°C Alkalihalobacillus wakoensis
0.062
-
1,2-naphthoquinone mutant enzyme W60A, at pH 8.5 and 25°C Alkalihalobacillus wakoensis
0.064
-
1,4-Naphthoquinone mutant enzyme W60A, at pH 8.5 and 25°C Alkalihalobacillus wakoensis
0.069
-
menadione wild type enzyme, at pH 8.5 and 25°C Alkalihalobacillus wakoensis
0.097
-
1,4-Naphthoquinone wild type enzyme, at pH 8.5 and 25°C Alkalihalobacillus wakoensis
0.115
-
Orange I wild type enzyme, at pH 8.5 and 25°C Alkalihalobacillus wakoensis
0.157
-
Orange I mutant enzyme W60A, at pH 8.5 and 25°C Alkalihalobacillus wakoensis

Organism

Organism UniProt Comment Textmining
Alkalihalobacillus wakoensis A0A455X8R2
-
-
Alkalihalobacillus wakoensis A01 A0A455X8R2
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-Sepharose column chromatography Alkalihalobacillus wakoensis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1,2-naphthoquinone + 2 NADH + 2 H+
-
Alkalihalobacillus wakoensis ?
-
?
1,2-naphthoquinone + 2 NADH + 2 H+
-
Alkalihalobacillus wakoensis A01 ?
-
?
1,4-naphthoquinone + 2 NADH + 2 H+
-
Alkalihalobacillus wakoensis ?
-
?
1,4-naphthoquinone + 2 NADH + 2 H+
-
Alkalihalobacillus wakoensis A01 ?
-
?
menadione + 2 NADH + 2 H+
-
Alkalihalobacillus wakoensis ?
-
?
menadione + 2 NADH + 2 H+
-
Alkalihalobacillus wakoensis A01 ?
-
?
additional information the enzyme shows a negligible ability to use molecular oxygen as an electron acceptor Alkalihalobacillus wakoensis ?
-
-
additional information the enzyme shows a negligible ability to use molecular oxygen as an electron acceptor Alkalihalobacillus wakoensis A01 ?
-
-
Orange I + 2 NADH + 2 H+
-
Alkalihalobacillus wakoensis ?
-
?
Orange I + 2 NADH + 2 H+
-
Alkalihalobacillus wakoensis A01 ?
-
?

Subunits

Subunits Comment Organism
homodimer
-
Alkalihalobacillus wakoensis

Synonyms

Synonyms Comment Organism
AzoA
-
Alkalihalobacillus wakoensis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
71
-
apparent melting temperature Alkalihalobacillus wakoensis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.85
-
Orange I wild type enzyme, at pH 8.5 and 25°C Alkalihalobacillus wakoensis
1.06
-
Orange I mutant enzyme W60A, at pH 8.5 and 25°C Alkalihalobacillus wakoensis
67
-
1,2-naphthoquinone mutant enzyme W60A, at pH 8.5 and 25°C Alkalihalobacillus wakoensis
111
-
1,2-naphthoquinone wild type enzyme, at pH 8.5 and 25°C Alkalihalobacillus wakoensis
136
-
menadione mutant enzyme W60A, at pH 8.5 and 25°C Alkalihalobacillus wakoensis
152
-
1,4-Naphthoquinone mutant enzyme W60A, at pH 8.5 and 25°C Alkalihalobacillus wakoensis
340
-
1,4-Naphthoquinone wild type enzyme, at pH 8.5 and 25°C Alkalihalobacillus wakoensis
387
-
menadione wild type enzyme, at pH 8.5 and 25°C Alkalihalobacillus wakoensis

Cofactor

Cofactor Comment Organism Structure
FMN
-
Alkalihalobacillus wakoensis
NADH the enzyme is strictly NADH-dependent Alkalihalobacillus wakoensis