Abstract
The purpose of this study was to screen and verify salt-enhancing peptides that can effectively reduce sodium consumption from Largemouth bass myosin through virtual hydrolysis, molecular simulation, and sensory evaluation. The human transmembrane channel-like 4 (TMC4) was constructed using Alphafold2, with 93.3 % of amino acids falling within allowed regions. A total of 19 peptides were predicted through virtual hydrolysis and screening. DAF, QIF, RPAL, and IPVM significantly enhanced the saltiness perception, and QIF exhibited the most pronounced effect in enhancing saltiness (P < 0.05). The residues Ala258, Ser546, Ser603, Phe259, Cys265, Glu539, Lys278 and Ser585 were identified as key binding sites. The TMC4-DAF complex achieved stability after 20, 000 ps, exhibiting an average RMSD value of 0.84 nm. DAF consistently displayed fluctuations at approximately 3.05 nm, and the number of hydrogen bonds varied between 3 and 5. These results suggested that Alphafold2 modelling can be used for predicting salt-enhancing peptides.
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