Fluorene degradation using immobilized Pleurotus ostreatus on various supports including polystyrene foam, Amberlite XAD-7, Amberlite XAD-2000, pumice particles and sand has been reported. These supports have yielded positive results with regard to performing metabolic functions, but the drawback of synthetic resins is that they are not eco-friendly. If natural supports could be used for cell immobilization for the degradation of fluorene, they would be both eco-friendly and perform metabolic functions. Therefore. This study attempts to design an immobilized P. ostreatus using pumice as a support specifically for the application of fluorene biodegradation in batch shaking systems. Laccase and MnP enzyme activities were monitored in these systems. The maximum fluorene degradation level was 89.16%. Batch degradation of fluorene in a packed bed reactor was also studied. The maximum fluorene degradation level was found to be 82.46%. Additionally, two different, synthetically concocted wastewater samples containing fluorene were studied in a recycled packed bed reactor in repeated mode. At the end of biodegradation process, the metabolites of the fluorene biodegradation produced by immobilized P. ostreatus were analyzed via GC-MS and FT-IR. It was concluded that the biotransformation of fluorene by immobilized P. ostreatus was achieved. (c) 2011 Elsevier Ltd. All rights reserved.