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Information on EC 1.7.1.17 - FMN-dependent NADH-azoreductase and Organism(s) Enterococcus faecalis and UniProt Accession Q831B2

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IUBMB Comments
Requires FMN. The enzyme catalyses the reductive cleavage of an azo bond in aromatic azo compounds to form the corresponding amines. Does not accept NADPH. cf. EC 1.7.1.6, azobenzene reductase.
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This record set is specific for:
Enterococcus faecalis
UNIPROT: Q831B2
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The taxonomic range for the selected organisms is: Enterococcus faecalis
The enzyme appears in selected viruses and cellular organisms
Synonyms
fmn-dependent nadh-azoreductase, indigo reductase, azoro, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
FMN-dependent-NADH azoreductase
-
-
SYSTEMATIC NAME
IUBMB Comments
N,N-dimethyl-1,4-phenylenediamine, anthranilate:NAD+ oxidoreductase
Requires FMN. The enzyme catalyses the reductive cleavage of an azo bond in aromatic azo compounds to form the corresponding amines. Does not accept NADPH. cf. EC 1.7.1.6, azobenzene reductase.
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4'-(dimethylamino)-azobenzene-2-carboxylic acid + 2 NADH + 2 H+
2-aminobenzoic acid + N,N-dimethyl-p-phenylenediamine + 2 NAD+
show the reaction diagram
i.e. methyl red
-
-
?
4'-(dimethylamino)-azobenzene-2-carboxylic acid + 2 NADPH + 2 H+
2-aminobenzoic acid + N,N-dimethyl-p-phenylenediamine + 2 NADP+
show the reaction diagram
i.e. methyl red
-
-
?
amaranth + 2 NADH + 2 H+
4-aminonaphthalene-1-sulfonic acid + 4-amino-3-hydroxynaphthalene-2,7-disulfonic acid + 2 NAD+
show the reaction diagram
-
-
-
?
orange G + 2 NADPH + 2 H+
aniline + 8-amino-7-hydroxynaphthalene-1,3-disulfonic acid + 2 NADP+
show the reaction diagram
-
-
-
?
orange II + 2 NADPH + 2 H+
4-aminobenzene-1-sulfonic acid + 1-aminonaphthalen-2-ol + 2 NADP+ + 2 NAD+
show the reaction diagram
-
-
-
?
ponceau BS + 2 NADH + 2 H+
? + 2 NAD+
show the reaction diagram
-
-
-
?
ponceau S + 2 NADH + 2 H+
? + 2 NAD+
show the reaction diagram
-
-
-
?
4'-(dimethylamino)-azobenzene-2-carboxylic acid + 2 NADH + 2 H+
2-aminobenzoic acid + N,N-dimethyl-p-phenylenediamine + 2 NAD+
show the reaction diagram
-
i.e. methyl red
-
-
?
4'-(dimethylamino)-azobenzene-2-carboxylic acid + 2 NADPH + 2 H+
2-aminobenzoic acid + N,N-dimethyl-p-phenylenediamine + 2 NADP+
show the reaction diagram
-
i.e. methyl red
-
-
?
amaranth + 2 NADH + 2 H+
4-aminonaphthalene-1-sulfonic acid + 4-amino-3-hydroxynaphthalene-2,7-disulfonic acid + 2 NAD+
show the reaction diagram
-
-
-
-
?
brilliant black + 2 NADH + 2 H+
?
show the reaction diagram
-
-
-
-
?
tartrazine + 2 NADH + 2 H+
?
show the reaction diagram
-
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
FMN
-
Km value 0.003 mM
NADPH
-
NADH is 180fold preferred over NADPH
additional information
no cofactor: NADPH
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.011 - 0.034
4'-(dimethylamino)-azobenzene-2-carboxylic acid
0.08 - 0.371
NADH
0.011
4'-(dimethylamino)-azobenzene-2-carboxylic acid
-
pH 7.2, 25°C
0.082
NADH
-
pH 7.2, 25°C
15
NADPH
-
pH 7.2, 25°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
137
substrate amaranth, pH 7.1, 23°C
139
substrate orange II, pH 7.1, 23°C
167
substrate ponceau S, pH 7.1, 23°C
181
substrate 4'-(dimethylamino)-azobenzene-2-carboxylic acid, pH 7.1, 23°C
0.626
-
cofactor NADPH, pH 7.0, 25°C
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
43000
gel filtration
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 23000, SDS-PAGE
dimer
2 * 23000, SDS-PAGE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
in complex with FMN, to 2.07 A resolution. The AzoA monomer shows a typical NAD(P)-binding Rossmann fold with a highly conserved FMN binding pocket. A salt bridge between Arg18 and Asp184 restricts the size of the flavin binding pocket such that only FMN can bind
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
N121Q
115% of wild-type activity
R21G/N121A
136% of wild-type activity
R21G/N121Q
171% of wild-type activity
R21K
80% of wild-type activity
R21K/N121A
41% of wild-type activity
R21K/N121Q
40% of wild-type activity
W105A
complete loss of both affinity for FMN and enzyme activity
W105F
31fold decrease in vmax value, Km value similar to wild-type
W105G
complete loss of both affinity for FMN and enzyme activity
W105H
8fold decrease in vmax value, Km value similar to wild-type
W105Q
68fold decrease in vmax value, Km value similar to wild-type
W105Y
22% reduction in vmax value
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Liu, Z.; Chen, H.; Shaw, N.; Hopper, S.; Chen, L.; Chen, S.; Cerniglia, C.; Wang, B.
Crystal structure of an aerobic FMN-dependent azoreductase (AzoA) from Enterococcus faecalis
Arch. Biochem. Biophys.
463
68-77
2007
Enterococcus faecalis (Q831B2)
Manually annotated by BRENDA team
Sun, J.; Kweon, O.; Jin, J.; He, G.; Li, X.; Cerniglia, C.; Chen, H.
Mutation network-based understanding of pleiotropic and epistatic mutational behavior of Enterococcus faecalis FMN-dependent azoreductase
Biochem. Biophys. Rep.
12
240-244
2017
Enterococcus faecalis (Q831B2), Enterococcus faecalis ATCC 700802 (Q831B2)
Manually annotated by BRENDA team
Punj, S.; John, G.
Purification and identification of an FMN-dependent NAD(P)H azoreductase from Enterococcus faecalis
Curr. Issues Mol. Biol.
11
59-66
2009
Enterococcus faecalis, Enterococcus faecalis ATCC 27274
Manually annotated by BRENDA team
Chen, H.; Xu, H.; Kweon, O.; Chen, S.; Cerniglia, C.
Functional role of Trp-105 of Enterococcus faecalis azoreductase (AzoA) as resolved by structural and mutational analysis
Microbiology
154
2659-2667
2008
Enterococcus faecalis (Q831B2), Enterococcus faecalis ATCC 700802 (Q831B2)
Manually annotated by BRENDA team
Chen, H.; Wang, R.; Cerniglia, C.
Molecular cloning, overexpression, purification, and characterization of an aerobic FMN-dependent azoreductase from Enterococcus faecalis
Protein Expr. Purif.
34
302-310
2004
Enterococcus faecalis (Q831B2), Enterococcus faecalis ATCC 19433 (Q831B2)
Manually annotated by BRENDA team
Zahran, S.; Ali-Tammam, M.; Hashem, A.; Aziz, R.; Ali, A.
Azoreductase activity of dye-decolorizing bacteria isolated from the human gut microbiota
Sci. Rep.
9
5508
2019
Bacillus cereus, Enterococcus avium, Enterococcus faecalis, Escherichia coli
Manually annotated by BRENDA team