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A reusable magnetic nanoparticlesbased cryogel optical biosensor was fabricated to determine of sucrose in sugar cane and sugar samples. In this design, magnetic nanoparticles were embedded in cryogel and then the multienzymes were immobilized. The magnetic nanoparticles in cryogel helped separate the immobilized multi-enzyme cryogel biosensor by applying the external magnetic field to reuse for the subsequent detection. The developed cryogel biosensor exhibited good selectivity for sucrose determination with the detection limit of 3 mM and can be reused up to 32 times with good repeatability (<5 % RSD). Furthermore, the proposed optical biosensor can also be utilized for sucrose analysis in sugar cane and sugar samples. [ABSTRACT FROM AUTHOR]