Effect of cellobiose on the myofibrillar protein denaturation induced by pH changes during freeze-thaw cycles

The aim of this study was to investigate myofibrillar protein (MFP) denaturation induced by pH changes during freeze-thaw (FT) cycles, and to propose an effective mitigation strategy. Owing to the selective crystallization of Na₂HPO₄·12H₂O and the consequent pH change, a pH change of 3.32 units was observed when the MFP solution were frozen. The surface hydrophobicity, particle size and confocal laser scanning microscopy showed that the protein molecules gradually unfolded and formed larger protein aggregation as the number of FT cycles increases. Additionally, protein degradation, secondary and tertiary structure alterations suggested that the FT cycle could disrupt structural integrity. The addition of cellobiose could maximize the inhibition of pH changes (decrease of ∼0.62 unit), no Na₂HPO₄·12H₂O crystallization was observed by X-ray diffraction. Cellobiose could minimize FT damage to myofibrillar protein, which was closest to the control. Thus, cellobiose can be used as a new and effective cryoprotectant.