Glucose oxidase is an oxidoreductase flavoprotein which has high range applications in food, pharmaceutical and in clinical chemistry. However, enzyme applications are prohibited due to some limitations of enzymes. Hence, there is a need to overcome these limitations by immobilization techniques. This study mainly focuses on magnetic nanoparticles-based immobilization of an enzyme that is successfully used because this method provides the greater stability and support to enzyme due to the high surface area and small size of nanoparticles. Different immobilization conditions were also optimized. Co-precipitation method was followed to synthesize the iron oxide nanoparticles and characterized using various spectrophotometric and analytical techniques like UV-Vis, SEM, FTIR, Zeta Sizer and XRD. Moreover, the role of magnetic and immobilized iron oxide nanoparticles as decolorizing agent was examined. Present results showed the successful synthesis of magnetic nanoparticles and confirmed by their characterization. Best immobilization conditions were achieved at temperature of 10°C, pH 6 and immobilization time of 2.5 hours while GOx/nanoparticles ratio of 3000U/g. From the present findings it is concluded that immobilization of glucose oxidase is an effective way for creating bioconjugates to develop economically feasible and high-performance catalysts with improved stability and reusability for a variety of applications in medical and food industries.