Representative azo dyes (Acid Blue 113 [AB113] and Direct Black 38 [DB38]) were treated in a single step with soybean peroxidase (SBP) and hydrogen peroxide (H2O2), or in two steps, zero-valent iron (Fe°) pretreatment followed SBP/H2O2. The purpose of this research was to compare both treatment processes and to determine which one was the optimal for degradation of each azo dye. For AB113, the preferred process was the single-step process, 1.0 mM AB113 required 2.5 mM H2O2, 1.5 U/mL SBP at pH 4.0 for ≥ 95% color and dye removal and 30% total organic carbon (TOC) removal. For DB38, due to the products formed after Fe° reduction, which are enzyme substrates (aniline and benzidine; two of four products) a two-step process was preferred, which allowed reduction in the required SBP and H2O2 concentrations by 5- and 2-fold, respectively, compared to a single-step treatment for ≥ 95% color, dye, and aniline/benzidine removal and 88% TOC removal.
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