Horseradish peroxidase(HRP) in organic solvent can be reactivated by evaporation. In order to measure the evaporation effect, the enzyme solutions were obtained by evaporation and dilution of organic solvent, respectively. Although two situations were thermodynamically identical, the activity from evaporation was higher than that from dilution. From the UV absorbance and the fluorescence intensity mesurements, it can be explained that reactivation of enzyme activity might be caused by reversible folding, and the enzyme obtained by evaporation was more refolded than that obtained by dilution.