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Summary: Recrystallisation often occurs in the freeze‐thaw cycles and results in poor food quality. This study was undertaken to analyse the effects of recrystallisation on the changes of protein conformation and moisture migration within red seabream fillets after the freeze‐thaw cycles. The UV second‐derivative spectroscopy, intrinsic fluorescence measurement and Raman spectroscopy were employed to evaluate the protein conformational changes. The Low‐field NMR spectra analysis was used to evaluate moisture migration. The results indicated that the freeze‐thaw cycles altered the secondary and tertiary structure of proteins. It was noticed that interactions between water and protein molecules were decreased gradually and eventually induced the thermal instability of the myosin molecules. The immobilised water content declined and the free water content increased. There were significant destructions in fillet muscle microstructure. These observations unambiguously reflect how the recrystallisations affect the protein conformation and moisture migration patterns of red seabream fillets during freeze‐thaw cycles. [ABSTRACT FROM AUTHOR]