A new biodesulphurization method has been considered using Pseudomonas aeruginosa supported on polyethylene (PE) for biodesulphurization (BDS) of thiophene as an aromatic sulphur model compound of crude oils. Also the biodegradation of thiophene has been modified in the presence of potassium hexacyanoferrate(III) as a terminal electron acceptor to approach the maximum biodesulphurization efficiency. The obtaining results according to UV-Spectrophotometry at 240 nm, 83.3% of thiophene at the primary concentration of 50 mg/dm3, pH = 7, by 0.5 g of biocatalyst in 37°C after 4 h of contact time has been removed. The bacterial cells exhibited a greater and faster biodegradation in the presence of potassium hexacyanoferrate(III) and 94.8% of thiophene has been removed after 3 h of contact time. Kinetic study predicted chemisorption of thiophene on the surface of the biocatalyst, as it followed the pseudo-second-order rate equation. Morphology and surface functional groups of the biocatalyst have been investigated by SEM and FT-IR, respectively.