The increase in flooding in Europe is linked to the presence of excessive moisture in many areas. Enzymes are a sensitive parameter of soil environment, therefore we have made an attempt to assess the impact of simulated flood conditions on their activity. We determined the activity of dehydrogenases and acid and alkaline phosphatase after 7 and 14 days of flooding the three Fluvisols with river water in controlled conditions. The obtained results indicate a significant influence of soil moisture on enzymatic activity and soil pH, which indicates that the soil environment is changing as a result of flooding.