Present study illustrates the significance of biocatalyst's reductive behaviour in the degradation of dye molecules using glucose as co-substrate. An anaerobic system was operated at a dye concentration of 50mg/l with an organic loading rate (OLR) of 1.36kg COD/m3-day. Decolourization and COD removal efficiencies were observed to be 42% and 48% respectively. Azo reductase (18.9U) and dehydrogenase enzyme (1.4μg/ml) activities showed increment with operation time. Anaerobic microenvironment showed dye reduction converting them into aromatic amines. The presence of mediators viz., cytochromes, quinines and Fe-S proteins depicted in the cyclic voltammetry profiles played a crucial role in transfer of electrons for the reduction of dye molecules. Bio-electro kinetic profiles obtained through Tafel analysis showed persistent reduction behaviour, which is in good correlation with dye degradation in the anaerobic microenvironment. © 2015 Elsevier Ltd.