The contribution of biotic and abiotic processes during azo dye reduction in anaerobic sludge

Frank P. Van Der Zee, Iemke A.E. Bisschops, Valérie G. Blanchard, Renske H.M. Bouwman, Gatze Lettinga, Jim A. Field

Research output: Contribution to journalArticlepeer-review

94 Scopus citations


Azo dye reduction results from a combination of biotic and abiotic processes during the anaerobic treatment of dye containing effluents. Biotic processes are due to enzymatic reactions whereas the chemical reaction is due to sulfide. In this research, the relative impact of the different azo dye reduction mechanisms was determined by investigating the reduction of Acid Orange 7 (AO7) and Reactive Red 2 (RR2) under different conditions. Reduction rates of two azo dyes were compared in batch assays over a range of sulphide concentrations in the presence of living or inactivated anaerobic granular sludge. Biological dye reduction followed zero order kinetics and chemical dye reduction followed second-order rate kinetics as a function of sulfide and dye concentration. Chemical reduction of the dyes was greatly stimulated in the presence of autoclaved sludge; whereas chemical dye reduction was not affected by living or γ-irradiated-sludge. Presumably redox-mediating enzyme cofactors released by cell lysis contributed to the stimulatory effect. This hypothesis was confirmed in assays evaluating the chemical reduction of AO7 utilizing riboflavin, representative of the heat stable redox-mediating moieties of common occurring flavin enzyme cofactors. Sulfate influenced dye reduction in accordance to biogenic sulfide formation from sulfate reduction. In assays lacking sulfur compounds, dye reduction only readily occurred in the presence of living granular sludge, demonstrating the importance of enzymatic mechanisms. Both chemical and biological mechanisms of dye reduction were greatly stimulated by the addition of the redox-mediating compound, anthraquinone-disulfonate. Based on an analysis of the kinetics and demonstration in lab-scale upward-flow anaerobic sludge bed reactors, the relative importance of chemical dye reduction mechanisms in high rate anaerobic bioreactors was shown to be small due to the high biomass levels in the reactors.

Original languageEnglish (US)
Pages (from-to)3098-3109
Number of pages12
JournalWater research
Issue number13
StatePublished - Jul 2003


  • Azo dye reduction
  • Granular sludge
  • Mechanism
  • Redox mediators
  • Sulfide

ASJC Scopus subject areas

  • Ecological Modeling
  • Water Science and Technology
  • Waste Management and Disposal
  • Pollution


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