Molecular modelling studies reveal cryoprotective mechanism of L-Proline during the frozen storage of shrimp (Litopenaeus vannamei).

• Incorporation of proline improved water-holding capacity of frozen shrimp. • Interactions between proline, water, and myosin were studied. • Proline improved the stability of shrimp myosin. • Proline may form a glassy state around shrimp myosin. • Proline inhibited ice crystal formation by forming hydrogen bonds with water. This study aimed to investigate the cryoprotective properties of proline (1% and 3% (w/v)) on shrimp. The cryoprotective mechanism was studied using physico-chemical experiments and molecular simulations. Proline had a notable positive impact on the thawing loss and texture of shrimp in comparison to the control. The denaturation of myosin in frozen shrimp was delayed by proline. Microscopy analysis demonstrated that proline effectively lowered the harm caused by ice crystals to shrimp muscle. Molecular simulations indicated that proline potentially exerted a cryoprotective effect primarily through the "water substitution" and "glassy state" hypotheses. Proline formed hydrogen bonds with myosin to replace the water molecules around myosin. Additionally, proline interacted with water molecules to form a glassy state, impeding the growth of ice crystals. Consequently, the stability of shrimp myosin was enhanced during freezing. In conclusion, proline demonstrated promise as an efficacious cryoprotectant for aquatic products. [ABSTRACT FROM AUTHOR]