This study was performed to investigate the effects of water molecules on ozone oxidation of acetaldehyde using a manganese oxide catalyst at room temperature. The catalytic ozone oxidation was conducted at different relative humidity (RH) conditions of 0%, 50%, and 80%. As the RH increased, both ozone and acetaldehyde removal efficiencies dropped due to competitive adsorption on the surface of the catalyst. At the highest RH of 80%, the oxidation reaction was severely retarded, and oxidation by-products such as acetic acid were formed and adsorbed on the surface. After the ozone oxidation of acetaldehyde, the regeneration of the catalyst using ozone alone was tested, and the further oxidation of accumulated organic compounds was investigated under the RH conditions of 0%, 50%, and 80%. When the highest relative humidity was introduced in the regeneration step, the ozonation reaction with the by-products adsorbed on the catalyst surface decreased due to the competitive reaction with water molecules. These findings revealed that, only when relative humidity was low to minimize the formation of by-products, the ozone oxidation of acetaldehyde using the manganese oxide catalyst at room temperature can be feasible as an effective control method.