Ice crystal-induced protein denaturation is the main cause of the deterioration of fish during frozen storage and transportation. In this study, the ultra-performance liquid chromatography − quadrupole − time of flight (UPLC-Q-TOF) technique was used to identify and screen tryptic peptides Ile-Glu-Glu-Leu-Glu-Glu-Leu-Glu-Ala-Glu-Arg (IEELEEELEAER) from large yellow croaker (Pseudosciaena crocea). The results were used study their cryoprotective effects on turbot fish meat during freeze–thaw cycles at different concentrations, and to investigate their anti-freezing mechanism. The results showed that the I-2.0 group effectively inhibiting the degeneration and structure changes of myofibrillar proteins after three freeze–thaw cycles, and the Ca2+-ATPase activity (1.65 μmolPi/mg/h), increased by 55.86% compared with that of the control group. Additionally, peptide IEELEEELEAER could provide antifreeze protection by binding to the surface of ice crystals and inhibiting their transformation. This peptide acts as a natural cryoprotectant and might be used for the cryogenic storage and transportation of fish products.