In our study, an electrospun cellulose acetate nanofibrous web (CA-NFW) was found to be quite effective in immobilizing bacterial cells. Here, decolorization of methylene blue (MB) dye in aqueous medium was achieved by using three types of bacteria (Aeromonas eucrenophila, Clavibacter michiganensis and Pseudomonas aeruginosa) immobilized on the CA-NFW. The decolorization time (0-48 h) and different MB dye concentrations (20-500 mg L-1) were studied to elucidate the maximum MB dye removal by the bacteria immobilized CA-NFWs. The effective dye decolorization was achieved within 24 hours and MB dye removal was similar to 95%. Interestingly, MB dye decolorization performance of bacteria immobilized CA-NFWs was quite close to that of free bacteria. We have also tested the reusability of bacteria immobilized NFWs after four cycles and similar to 45% of the dye decolorization capacity was obtained at the end of the 4th cycle. These results are quite promising and therefore suggest that bacteria immobilized electrospun NFWs could be quite applicable for the decolorization of dyes in wastewater due to their versatility and reusability.