厚壳贻贝乙醇脱氢酶激活肽的纯化、鉴定及其与ADH的相互作用OA北大核心CHSSCDCSTPCD

[Objective]To develop the alcohol dehydrogenase(ADH)activating peptide from Mytilus coruscus and to investigate the molecular mechanism of its binding with ADH.[Method]Using the in vitro ADH activation rate,M.coruscus peptides(MCP)with molecular less than 1 ku was separated and purified by gel column chromatography and reversed phase liquid chromatography(RP-LC).The peptides with high ADH activation were identified by liquid chromatography tandem mass spectrometry(LC-MS/MS),and the interaction between ADH was explored by molecular docking technology.[Result]The active fraction Y-I-I was separated by chromatography from MCP,and three active peptide sequences PPLYE,PPLYQ and APPLYQ with lower docking binding energy with ADH were identified and selected.Among them,the active peptide PPLYE had the best effect in ADH activation in vitro,reaching 77.35%at 2.0 mg/mL in a dose-dependent manner.Molecular docking results showed that PPLYE could be docked to the hydrophobic cavity near the active center of ADH,and form a stable complex with the receptor protein ADH through hydrophobic interaction and hydrogen bonding,with a lowest binding energy of-35.58 kJ/mol.[Conclusion]PPLYE,as the best ADH activating peptide can interact with ADH through hydrophobic interaction and hydrogen bonds to form a stable compound and increase ADH activity.This study provides a basis for the exploitation and utilization of M.coruscus resources and the development of ADH activating peptide.