Glucose oxidase (EC 126.96.36.199) sensors that have been developed and widely used for glucose monitoring have generally relied on electrochemical principle. In this study, the potential use of colorimetric method for glucose detection utilizing glucose oxidase-magnetic cellulose nanocrystals (CNCs) is explored. Magnetic cellulose nanocrystals (magnetic CNCs) were fabricated using iron oxide nanoparticles (IONPs) and cellulose nanocrystals (CNCs) via electrostatic self-assembly technique. Glucose oxidase was successfully immobilized on magnetic CNCs using carbodiimide-coupling reaction. About 33% of GOx was successfully attached on magnetic CNCs, and the affinity of GOx-magnetic CNCs to glucose molecules was slightly higher than free enzymes. Furthermore, immobilization does not affect the specificity of GOx-magnetic CNCs towards glucose and can detect glucose from 0.25 mM to 2.5 mM. Apart from that, GOx-magnetic CNCs stored at 4 °C for 4 weeks retained 70% of its initial activity and can be recycled for at least ten consecutive cycles.