In this report, morphology and fiber diameter differences of electrospun polysulfone (PSU) fibers on bacterial immobilization and bioremediation performance were evaluated. PSU fibers were produced with aligned or randomly oriented morphologies, and PSU fibers with thinner and thicker diameters were also produced. PSU fibers were utilized as carrier matrices for bacterial integration and the sample showing highest bacterial immobilization was tested for bioremediation of ammonium and methylene blue dye in water. It was found that randomly oriented and thinner PSU fibers are the optimal system for bacterial immobilization, hence bioremediation studies were performed with this sample. The results demonstrated that bacteria immobilized PSU fibers are promising candidates for simultaneous removal of ammonium and methylene blue dye, and they have a potential to be used in remediation of water systems.