Cholesterol oxidase enzyme well known as oxidoreductase enzyme which able to catalyze the transfer of electron from one molecule (the oxidant) to another molecule (the reductant). The kinetic model for cholesterol oxidation by cholesterol oxidase has been demonstrated using first order irreversible reaction with Euler’s method. For this kinetic study, experimental data was obtained from several literatures. The affecting factors of kinetic rate constant in this study were solvent addition, stabilizer concentration, granule concentration, temperature and immobilized enzyme. In the oxidation reaction, there were some variables condition based on the experimental data, CA-1, CA-2, CA-3, respectively. The modelling result showed that the first order reaction was able to modelling the cholesterol oxidation with high accuracy. This kinetic study is useful for further developing of application for biosensor based enzyme as catalyzer in the oxidation reaction. Moreover, kinetic study is also useful for choosing the operating factor for cholesterol oxidation.