An important portion of organic matter and colorants still remain in the biologically treated distillery wastewater, leaving the dark brown and odorous downstream with the heavy loading of chemical oxygen demand and the potential of forming disinfection byproducts. However, those bio-recalcitrant colorants have not been clearly recognized. The current study investigated the features of the bio-refractory organic matter and colorants in a typical distillery effluent, cassava distillery wastewater; special attention was paid to their change and behaviors in the coagulation treatment following the bio-processes. The wastewater analyses denoted that the fraction of high molecular weight (1-50 kDa and >50 kDa) became predominant after the anaerobic-aerobic processes. Importantly, the lignin breakdown products, melanoidins and lignin phenols were confirmed to be the leading colored components, according to the parallel factor analysis of fluorescence excitation-emission matrixes results. Compared with lignin phenols, the former two types of colorants exhibited stronger bio-refractory activity and resulted in smaller color reduction after the aerobic treatment. Neither advanced oxidation nor adsorption could perform efficiently as post-treatment for decolorization in this study. Nevertheless, high removal of color and dissolved organic matter (∼94.0% and ∼78.3%, respectively) could be achieved by the FeCl-involved coagulation under the optimal conditions. The ferric coagulant was found to preferably interact with the aromatic compounds (such as lignin derivatives) and melanoidins via either surface complexation or electric charge neutralization, or both. The findings presented herein might provide an insight into the evaluation of bio-refractory organic colorants and the Fe(III)-involved decolorization mechanisms of ethanol production wastewaters.