The aim of this study is to comparatively investigate the decolorization of Reactive Blue 171 (RB 171) by using three different bacterial strains as Bacillus megaterium A1 (A1), Gordonia sp. MC-D1 (D1), and Bacillus pumilus D3 (D3) and also a white rot fungus (Pleurotus ostreatus). All tested bacteria were incubated with RB 171 dye at 150 mg/L concentration for 24-72 h under static or agitated (150 rpm) conditions while the fungus was incubated with 150 mg/L of the dye under the same conditions for 3-24 h. The highest bacterial decolorization values were obtained after 72 h of incubation under static conditions, and the maximum decolorization rates were detected as 84, 83 and 75% for A1, D1, and D3, respectively. However, the color of RB 171 dye was removed at 93% rate by Pleurotus ostreatus under static conditions after 24 h of incubation. Similar results were also obtained from the agitated studies of the tested fungal and bacterial strains except A1. The maximum Birhanlı et al. (2021) ADYU J SCI, 11(2), 350-361 351 decolorization values obtained with A1, D1, and D3 at 150 rpm, 72 h were 30, 88 and 89%, respectively. The highest decolorization activities of Pleurotus ostreatus were 93% for both static and agitated (150 rpm) conditions. In addition, zymogram analyzes of the fungal culture fluids obtained from SBM, and SBM containing 150 mg/L RB 171 were also performed to detect the presence of laccase.