Changes in proteins and the taste-active substance content of oysters sterilized with ozonated water were investigated in this study. Shelled fresh oyster meat was treated with 4 mg•L-1 ozonated water for 2 min, and the changes in fluorescence and infrared spectra, particle size, solubility, turbidity, and the taste-active nucleotide and amino acid contents of oyster meat following sterilization were analyzed during storage at 0 ℃. Ozonated water sterilization led to protein aggregation in oyster meat, as evidenced by increased particle size and turbidity. The content of active sulfhydryl groups in oysters was significantly decreased (P<0.05); however, the degree of aggregation in oyster meat during the late stage of frozen storage was improved. No significant changes were observed in the secondary and tertiary structures of oyster myofibrillar proteins (P>0.05). Inosine 5′-monophosphate (IMP) was the primary taste-active nucleotide of the oysters. Ozonated water sterilization increased IMP content (P<0.05) and decreased that of guanosine 5'-monophosphate (P<0.05) in oyster meat. The taste-active nucleotide content of all groups significantly decreased (P<0.05) during the late stage of storage. Six taste-active amino acids were detected in oyster meat, accounting for 33.48% of the total amino acid content. Following ozonated water sterilization, the proportion of taste-active amino acids decreased to 30.18%. In conclusion, this study found no significant changes in the myofibrillar protein structure of ozonated water-sterilized oyster meat during frozen storage. Protein stability during the late stage of frozen storage was enhanced, while the content of taste-active substances was reduced. The results of this study can serve as a reference for the application of ozonated water in the preservation of the freshness of aquaculture products.