In today's world, there is a growing gap between energy demand and limited energy sources such as fossil fuels. As a result, renewable energy sources such as hydrogen are increasingly being researched. Hydrogen is considered one of the most promising energy sources of the future, as it has a high energy yield. Hydrogen produced from biological sources is known as biohydrogen and is one of the most interesting renewable energy sources today. One way of producing biohydrogen is the enzymatic conversion of glucose into gluconic acid with the regeneration of coenzymes and the parallel production of molecular hydrogen. In this study, glucose oxidation, catalyzed by the enzyme glucose dehydrogenase (GDH) with the coenzyme NAD+ was carried out in a batch reactor. In the first part of the study, the characterization of the enzyme GDH was performed by determining the optimal temperature and pH for the reaction and selecting the best buffer. The results showed that the optimal temperature for the reaction is 40°C, pH 7, and the buffer is 0.1 mol/L TRIS-HCl pH 7. In the second part of the study, the reaction kinetics of glucose oxidation was investigated using the initial rate method, and the kinetic parameters were estimated. The kinetics of glucose oxidation catalyzed by GDH is described by two-substrate Michaelis-Menten kinetics with substrate inhibition and competitive product inhibition. A suitable reactor model was developed for the reaction carried out in the batch reactor, consisting of a kinetic model and mass balances for all components of the reaction mixture. The validation of the model was evaluated using two sets of independent experiments.