Immobilized quinones exhibit good catalytic performance in the biodecolorization of azo dyes. However, in practical activated sludge systems, little is known about the effect of azo dye concentration on microbial communities in the presence of immobilized quinones. 454 Pyrosequencing was used to investigate structural changes and to determine the key microorganisms involved in Reactive Red X-3B decolorization in the presence of anthraquinone-2-sulfonate immobilized on polyurethane foam (AQS-PUF). Our results show that the AQS-PUF-supplemented system exhibited better stability and decolorization performance during a 30-day run than polyurethane-foam-only (PUF-supplemented) and control systems. Analysis of pyrosequencing data showed that the AQS-PUF-supplemented system had the highest bacterial diversity, followed by the control and PUF-supplemented systems during decolorization. Reactive Red X-3B and AQS-PUF significantly influenced bacterial communities at the class level: Erysipelotrichia and the most dominant Deltaproteobacteria showed significant positive correlations with Reactive Red X-3B, while unclassified Firmicutes were found to be significantly correlated with AQS-PUF. At the genus level, Desulfomicrobium, which represents 8-44 % of the total population, displayed a significant positive correlation with Reactive Red X-3B. Some bacteria, including Desulfovibrio, Shewanella, and Clostridium with relative abundances of less than 6 %, were positively correlated with AQS-PUF. These findings provide a novel insight into the changes that occur in the bacterial community during immobilized AQS-mediated decolorization. Less abundant quinone-reducing bacteria play important roles in accelerating the effect of AQS-PUF on biodecolorization.