This study investigated the role of oxidative stress in the mitochondrial apoptotic pathways and structural protein degradation of fish during postmortem storage by measuring oxidative stress levels, mitochondrial antioxidant enzyme activity, mitochondrial dysfunction, apoptotic factors, and structural protein degradation (n = 3). The results revealed that reactive oxygen species (ROS) increased gradually within the first 12 h and then decreased (p < 0.05) in mitochondria. Lipid peroxidation was increased, and superoxide dismutase, catalase, and glutathione peroxidase activities were decreased in mitochondria (p < 0.05). Furthermore, oxidative stress induced mitochondrial membrane opening, mitochondrial swelling, as well as the depolarization of mitochondrial potential. This led to an increase in the release of cytochrome c from mitochondria and caspase-3 activation. Ultimately, oxidative stress promoted small protein degradation (troponin-T and desmin) and induced myofibril susceptibility to proteolysis. These observations confirmed that oxidative stress mediated the activation of mitochondrial apoptotic factors-promoted protein degradation, initiating the deterioration of fish muscle through the mitochondrial apoptotic pathway.

The effects of changes in the malondialdehyde (MDA) content on the quality of fish during the low-temperature storage period are unclear. Therefore, the effects of the MDA content on Coregonus peled quality and protein changes were investigated following storage under refrigeration (4 °C) and super chilling (−3 °C) for 15 days (d). The results showed that the MDA content continued to increase during storage and that the highest content was produced at 1.42 mg/kg during refrigeration. The fillet pH, drip loss, texture (hardness and elasticity), and myofibril fragmentation index deteriorated significantly during the storage period. Increased oxidation of the myofibrillar protein (MP) was observed in the 15 d storage period, and the MP carbonyl content was 1.19 times higher under refrigeration than in super chilling, while the protein α-helix structure decreased by 12.48% and 12.20% under refrigeration and super chilling, respectively. Electropherograms also showed that myosin degradation was particularly severe in the refrigeration storage period of 15 d. Overall, the MDA formed at the refrigeration and super chilling storage temperatures could promote structural changes in, and the oxidative degradation of, proteins to different degrees, leading to the deterioration of the fillet quality. This study provides a scientific basis for investigating the relationship between fish quality and changes in the MDA content during low-temperature storage.

As the most valuable protein in the sperm of testes tissues of Coregonus peled, Coregonus peled protamine (CPP) had a natural antibacterial and antiseptic effect, but its physicochemical properties and functions are easily affected by the ultrasound-assisted extraction process. In this study, ultrasound-assisted extraction of CPP was used to investigate the effects of different ultrasonic intensities (0, 3.03, 6.07, 9.10, 12.13, and 15.16 W/cm²) on the structural and functional properties of CPP. The results showed that at moderate ultrasonic intensity (9.10 W/cm²), the protein was the most successful, as it was subjected to cavitation shear and microjet by ultrasound, which resulted in changes in protein structure, moderate unfolding of peptide chains, and changes in the secondary and tertiary structures of CPP. SEM images confirmed the changes in the microstructure of CPP. Ultrasound oxidized the proteins to varying degrees with the highest sulfhydryl and carbonyl and surface hydrophobicity content at an ultrasonic intensity of 9.10 W/cm². At the same time, the solubility, antimicrobial activity, and heparin binding of CPP were affected. It is worth mentioning that the ultrasonicated CPP exhibited a stronger heparin-binding capacity compared to the non-ultrasonicated CPP. In conclusion, 9.10 W/cm² was determined as the optimal ultrasonic intensity parameter for this study. In conclusion, the incorporation of appropriate ultrasonic intensity in the acidic extraction process could help to improve the functional properties of CPP, and ultrasound-assisted protein extraction has emerged as a reliable technique capable of modifying the structure and function of CPP.

The most frequently occurring protein modification in fish postmortem is oxidization, which further affects meat quality through multiple biochemical pathways. To investigate how hydroxyl radicals affect the structure of cathepsin H and its ability to break down myofibrillar proteins in Coregonus peled, cathepsin H was oxidized with 0, 0.1, 0.5, 1, 5, and 10 mM H₂O₂ and subsequently incubated with isolated myofibrillar proteins. The results showed that as the H₂O₂ concentration increased, the carbonyl and sulfhydryl contents of cathepsin H significantly increased and decreased, respectively. There were noticeable changes in the α-helix structures and a gradual reduction in UV absorbance and fluorescence intensity, indicating that oxidation can induce the cross-linking and aggregation of cathepsin H. These structural changes further reduced the activity of cathepsin H, reaching its lowest at 10 mM H₂O₂, which was 53.63% of the activity at 0 mM H₂O₂. Moreover, desmin and troponin-T all degraded at faster rates when cathepsin H and myofibrillar proteins were oxidized concurrently as opposed to when cathepsin H was oxidized alone. These findings provide vital insights into the interaction mechanism between oxidation, cathepsin H, as well as myofibrillar protein degradation, laying a groundwork for understanding the molecular mechanisms underlying changes in fish meat quality after slaughter and during processing.

In this study, cathepsin D was oxidized in vitro with different concentrations of H₂O₂, and the activity, structure, and extent of myofibrillar protein degradation by oxidized cathepsin D were evaluated. The sulfhydryl content of cathepsin D decreased to 9.20% after oxidation, while the carbonyl content increased to 100.06%. The β-sheet in the secondary structure altered due to oxidation as well. The changes in the intrinsic fluorescence and UV absorption spectra indicated that oxidation could cause swelling and aggregation of cathepsin D molecules. The structure of cathepsin D could change its activity, and the activity was highest under 1 mM H₂O₂. Cathepsin D could degrade myofibrillar proteins in different treatment groups, and the degree of degradation is various. Therefore, this study could provide a scientific basis for the mechanism of interaction among hydroxyl radical oxidation, cathepsin D, and MP degradation.