The kinetics of Glucose oxidase (GOD) and Horseradish peroxidase (HRP) on a transparent Ion Jelly biopolymer has been investigated using a colorimetric assay of H2O2 with phenol-4-sulfonic acid and 4-aminoantipyrine, as color-generating precursors. The activity of GOD and HRP on the Ion Jelly biopolymer solid disk formed in the bottom of the well ELISA plate showed to be respectively 16 times and 13 times lower than in free sodium phosphate buffer. The storage stability at 4ºC of the both immobilized enzymes showed that GOD can retain up to 70% of the initial activity after two weeks, where HRP retained 91% of its initial activity. The drop cast deposition of the transparent Ion Jelly biopolymer containing the two enzymes (GOD, HRP) and the color-generating precursors on filter paper allowed to demonstrate their applicability on colorimetric glucose detection.