Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.7.1.6 extracted from

  • Ito, K.; Nakanishi, M.; Lee, W.C.; Zhi, Y.; Sasaki, H.; Zenno, S.; Saigo, K.; Kitade, Y.; Tanokura, M.
    Expansion of substrate specificity and catalytic mechanism of azoreductase by X-ray crystallography and site-directed mutagenesis (2008), J. Biol. Chem., 283, 13889-13896.
    View publication on PubMed

Application

Application Comment Organism
industry industrial use of the enzyme in the biodegradation process of azo compounds Escherichia coli

Cloned(Commentary)

Cloned (Comment) Organism
into the vector pET-22b for expression in Escherichia coli BL21DE3 cells Escherichia coli

Crystallization (Commentary)

Crystallization (Comment) Organism
the structures of oxidized and reduced AzoR, and in complex with the inhibitor 3,3'-methylene-bis(4-hydroxycoumarin) are determined at resolutions from 1.4 to 2.3 A Escherichia coli

Protein Variants

Protein Variants Comment Organism
F162A Phe-162 is chosen because it is predicted to participate in the substrate binding on top of the isoalloxazine ring, as observed in the AzoR-inhibitor structure Escherichia coli
R59A the results indicate that Arg-59 decides the substrate specificity of AzoR Escherichia coli
Y120A Tyr-120 is chosen because it is predicted to participate in the substrate binding on top of the isoalloxazine ring, as observed in the AzoR-inhibitor structure Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
3,3'-methylene-bis(4-hydroxycoumarin) dicoumarol Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0102
-
4'-dimethylaminoazobenzene-2-carboxylic acid mutant Y120A Escherichia coli
0.0419
-
4'-dimethylaminoazobenzene-2-carboxylic acid wild-type enzyme Escherichia coli
0.0706
-
NADH wild-type enzyme Escherichia coli
0.0849
-
4'-dimethylaminoazobenzene-2-carboxylic acid mutant R59A Escherichia coli
0.1006
-
p-methyl red mutant R59A Escherichia coli
0.101
-
4'-dimethylaminoazobenzene-2-carboxylic acid mutant F162A Escherichia coli
0.23
-
p-methyl red mutant Y120A Escherichia coli
0.266
-
p-methyl red wild-type enzyme Escherichia coli
0.403
-
NADH mutant R59A Escherichia coli
0.637
-
NADH mutant Y120A Escherichia coli
0.739
-
NADH mutant F162A Escherichia coli
0.867
-
p-methyl red mutant F162A Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
23000
-
-
Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli P41407
-
-

Purification (Commentary)

Purification (Comment) Organism
His-tagged proteins are purified by gravity-flow chromatography using nickel-nitrilotriacetic acid-agarose Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-methyl-1,4-naphthoquinone + NADH + H+ menadione, vitamin K3 Escherichia coli ?
-
?
4'-diethylaminoazobenzene-2-carboxylic acid + NADH + H+ ethyl red Escherichia coli ?
-
?
4'-dimethylaminoazobenzene-2-carboxylic acid + NADH + H+ methyl red Escherichia coli 2-aminobenzoic acid + N,N'-dimethyl-p-phenylenediamine + NAD+
-
?
p-methyl red + NADH + H+
-
Escherichia coli ?
-
?

Subunits

Subunits Comment Organism
homodimer 2 * 23000 Escherichia coli

Synonyms

Synonyms Comment Organism
AzoR
-
Escherichia coli
azoreductase
-
Escherichia coli

Cofactor

Cofactor Comment Organism Structure
FMN
-
Escherichia coli
NADH
-
Escherichia coli