Purification of two azoreductases from Escherichia coli K12

Two azoreductases (I and II) were purified to homogeneity from extracts of E. coli K12. Azoreductase I was a dimer of two identical subunits of molecular weight 28000 whereas azoreductase II was a monomer of 12,000 molecular weight. Both NADH and NADPH functioned as electron donors for the azoreductases. Azoreductase I and II used Ponceau SX, Tartrazine, Amaranth and Orange II as substrates. Ponceau SX was the best substrate for both the enzymes. However, azoreductase II utilized tartrazine, amaranth and orange II less efficiently than azoreductase I.