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1.7.1.17: FMN-dependent NADH-azoreductase

This is an abbreviated version!
For detailed information about FMN-dependent NADH-azoreductase, go to the full flat file.

Reaction

anthranilate
+
N,N-dimethyl-1,4-phenylenediamine
+ 2 NAD+ =
2-(4-dimethylaminophenyl)diazenylbenzoate
+ 2 NADH + 2 H+

Synonyms

acpD, AZO, azo-dye reductase, AzoA, AzoR, AzoR1, azoreductase, AzoRo, AzrA, AzrG, flavin-containing oxygen-insensitive azoreductase, FMN dependent azoreductase, FMN-dependent methyl red reductase, FMN-dependent NADH-azo compound oxidoreductase, FMN-dependent NADH-azo reductase, FMN-dependent NADH-azoreductase, FMN-dependent-NADH azoreductase, indigo reductase, NADH-azoreductase

ECTree

     1 Oxidoreductases
         1.7 Acting on other nitrogenous compounds as donors
             1.7.1 With NAD+ or NADP+ as acceptor
                1.7.1.17 FMN-dependent NADH-azoreductase

Purification

Purification on EC 1.7.1.17 - FMN-dependent NADH-azoreductase

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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
native enzyme 4500fold by anion exchange chromatography, Blue Sepharose affinity chromtography, gel filtration, hydroxyapatite chromatography, again anion exchange chromatography and hydrophobic interaction chromatography. Recombinant enzyme AcpD from Escherichia coli, strain JM109 by anion exchange chromatography, Blue Sepharose affinity chromtography, gel filtration, and chromatography on GIGAPITE resin, the purified gene product exhibits activity corresponding to that of the native azoreductase
Ni-NTA column chromatography
Ni-Sepharose column chromatography
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
recombinant His-tagged enzyme from Escherichia coli strain XL1-Blue by nickel affinity chromatography
recombinant His-tagged wild-type and mutant AzoR enzymes from Escherichia coli BL21(DE3) by nickel affinity chromatography
recombinant His-tagged wild-type and mutant enzyme from Escherichia coli strain BL21(DE3) by cobalt affinity chromatography, dialysis, and ultrafiltration