Literature summary for 1.7.1.17 extracted from

Ito, K.; Nakanishi, M.; Lee, W.-C.; Sasaki, H.; Zenno,S.; Saigo, K.; Kitade, Y.; Tanokura, M.
Crystallization and preliminary X-ray analysis of AzoR (azoreductase) from Escherichia coli (2005), Acta Crystallogr. Sect. F, 61, 399-402 .

No PubMed abstract available

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme, sitting drop vapour-diffusion method, mixing 0.015 ml of 23 mg/ml protein in 10 mM Tris-HCl, pH 8.0, and 1 mM FMN, with an equal volume of reservoir solution containing 200 mM MgCl2, 30% v/v 2-propanol, and 100 mM HEPES, pH 7.5, equilibration over 0.5 ml reservoir solution, one week, 15°C, method optimization, crystal soaking in heavy metal solution with K2PtCl4, X-ray diffraction structure determination and analysis at 1.8-2.5 A resolution. FMN is tightly bound to the protein moiety, and this interaction is essential for the crystallization of AzoR	Escherichia coli

Crystallization (Comment)

Organism

purified recombinant enzyme, sitting drop vapour-diffusion method, mixing 0.015 ml of 23 mg/ml protein in 10 mM Tris-HCl, pH 8.0, and 1 mM FMN, with an equal volume of reservoir solution containing 200 mM MgCl2, 30% v/v 2-propanol, and 100 mM HEPES, pH 7.5, equilibration over 0.5 ml reservoir solution, one week, 15°C, method optimization, crystal soaking in heavy metal solution with K2PtCl4, X-ray diffraction structure determination and analysis at 1.8-2.5 A resolution. FMN is tightly bound to the protein moiety, and this interaction is essential for the crystallization of AzoR

Escherichia coli

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
cytoplasm	-	Escherichia coli	5737	-

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
2-(4-dimethylaminophenyl)diazenylbenzoate + 2 NADH + 2 H+	Escherichia coli	i.e. azo dye methyl red	anthranilate + N,N-dimethyl-1,4-phenylenediamine + 2 NAD+	-	?

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	P41407	-	-

Reaction

Reaction	Comment	Organism	Reaction ID
anthranilate + N,N-dimethyl-1,4-phenylenediamine + 2 NAD+ = 2-(4-dimethylaminophenyl)diazenylbenzoate + 2 NADH + 2 H+	ping-pong reaction mechanism	Escherichia coli	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
2-(4-dimethylaminophenyl)diazenylbenzoate + 2 NADH + 2 H+	i.e. azo dye methyl red	Escherichia coli	anthranilate + N,N-dimethyl-1,4-phenylenediamine + 2 NAD+	-	?
additional information	AzoR utilizes NADH but not NADPH as an electron donor and binds FMN as a flavin cofactor	Escherichia coli	?	-	?

Synonyms

Synonyms	Comment	Organism
AzoR	-	Escherichia coli
azoreductase	-	Escherichia coli
FMN-dependent NADH-azo compound oxidoreductase	-	Escherichia coli
FMN-dependent NADH-azo reductase	-	Escherichia coli

Cofactor

Cofactor	Comment	Organism
FMN	FMN is tightly bound to the protein moiety, and this interaction is essential for the crystallization of AzoR	Escherichia coli
additional information	AzoR utilizes NADH but not NADPH as an electron donor and binds FMN as a flavin cofactor	Escherichia coli
NADH	-	Escherichia coli

pI Value

Organism	Comment	pI Value Maximum	pI Value
Escherichia coli	sequence calculation	-	4.92