Aims: This study determined the optimum temperature for cell immobilization, the optimum time of fructose production by immobilized cell, and immobilized cell stability against repeated use in fructose production. Methodology and results: Research on cell immobilization of Streptomyces griseus and the variant have been done. The S. griseus variant was resulted from UV mutation. The variant was able to produce fructose as hydrolysis product 3 times as much after 30 min. Heating was done at 50, 60, 70, 80 and 90 °C. The fructose production was performed at intervals of 4 h for 32 h. The results showed that the optimum cell immobillization temperature of S. griseus and its variant was 80 °C. The optimum time of fructose production by immobilizing cell of S. griseus was 28 h and its variant cells was 24 h. Immobilized cells of S. griseus can be reused for 6 × 28 h to produce fructose compared to variants cells was 5 × 24 h, respectively. Conclusion, significance and impact of study This study reported that immobilized cells of S. griseus can be reused and its variant were highly advantageous in the production of fructose. The amount of fructose production was increased as compared to the conventional method and the cost of production could be reduced as well.