Cryoprotective saccharides are widely accepted additives of frozen seafood that reduce thawing loss,maintain texture,and slow down protein denaturation.The present study aimed to investigate the roles of trehalose and alginate oligosaccharides on cryoprotection of frozen shrimp,primarily focusing on the interactions between myosin and saccharide molecules using molecular dynamics (MD) simulation analysis.The results indicated that the incorporation of trehalose and alginate oligosaccharides markedly improved the capacities for thawing and cooking loss of frozen shrimp,with respective values decreasing to 6.02%,8.14%,and 5.99%,8.19% after 9 weeks of storage,which were significantly lower than that of fresh water treatment (9.75% and 15.09%).Our assumption was that water replacement played a leading role in cryoprotection,as shown in previous experimental results and reports.Furthermore,homology modeling and MD simulations confirmed that trehalose and alginate oligosaccharides substituted the water molecules around the myosin surface by forming hydrogen bonds with polar residues of amino acids,thereby stabilizing the structures in the absence of water during frozen storage.These conditions affected the flexibility of particular amino acid residues,enhanced the residue cross correlations within the two chains of myosin,and also increased the total interaction energy between myosin and water/saccharide molecules,thereby leading to an increase in protein stability.Finally,by comparing the experimental results to that of MD simulation,significant positive correlations were observed between saccharides and the stabilization of myosin in shrimp muscle.The findings of the present study may help in better understanding the cryoprotective mechanisms of saccharides in frozen shrimp,and the two saccharides may be potentially used as alternative additives in seafood to maintain better quality during frozen storage.