Bioethanol Mill Wastewater Purification by Combination of Coagulation-Flocculation and Microbial Treatment of Trametes versicolor INACC F200

Black liquor is generated from the pretreatment process of biomass-based bioethanol production and due its environmental impact, should be treated effectively before discharged to the water body. Chemical treatment using coagulation-flocculation method was commonly used for wastewater treatment. In the case of black liquor, chemical treatment is often insufficient and further treatment was needed to degrade lignin in order to reduce its black coloration. This present study investigated the two-step treatment to decolorize black liquor using chemical coagulation-flocculation and biological treatment using white-rot fungus Trametes versicolor INACC F200. The biological treatment was optimized by applying a response surface methodology (RSM) of the utilization of CuSO₄ concentration, Tween 80 concentration, and agitation. Furthermore, lignin degradation was also confirmed using FTIR and LC-MS. Initial chemical treatment using ferrous sulfate and polyacrylamide as coagulant-flocculant with a ratio of 3:3, resulted in black liquor decolorization at 80.9% and reduced the COD up to 90.77%. A full quadratic stepwise model was utilized with CuSO₄ inducer, Tween 80 mediator, and agitation speed as the independent variables. Optimum decolorization of 96.188% was predicted when using 2 mM CuSO₄, 2% Tween 80, and an agitation speed of 150 rpm. The highest enzyme activity during the decolorization process was lignin peroxidase (LiP). FT-IR and LC-MS profile showed that lignin-associated bond was eliminated and the molecular weight of lignin was decreased after the treatment. This study concludes the effective decolorization and delignification of black liquor by the two-step chemical and biological treatment.