Alcohol hydydrogenase (ADH) is a key enzyme that influences alcohol metabolism in vivo. Our previous study showed that chicken-derived tripeptide KPC could stabilize ADH in a dose-dependant manner. In this study, possible mechanism underlying how KPC could stabilize ADH was further investigated. Fluorescence quenching data showed that KPC induced a dynamic fluorescence quenching with a quenching rate constant value of 1.074 × 10¹⁰ M⁻¹s⁻¹, indicating a conformational change. Circular dichroism further suggested a possible transformation from α-helix (14.03%–13.90%) to random coil (31.98–33.04%) in ADH structure as affected by KPC (5 mmol/L). Molecular docking analysis predicted that KPC may bind to ADH through hydrophobic interaction and hydrogen bonds. Validation of ADH stabilizing activity of fragment peptides and amino acids from KPC implied the vital role of Cys residue to the bio-activity of KPC. These results indicated that KPC may stabilize ADH through peptide-enzyme interactions, and protect ADH against oxidative modification. Besides, a favorable absorption, distribution, metabolism, excretion, and toxicity (ADMET) profile of KPC was obtained using in silico assays. Overall, this study provided a comprehensive understanding on the interaction mechanism between KPC and ADH, and illustrated the potential applicability of KPC as drug or functional food ingredient.

酒精加氢酶（ADH）是影响体内酒精代谢的关键酶。我们之前的研究表明，鸡源性三肽 KPC 可以以剂量依赖的方式稳定 ADH。在这项研究中，进一步研究了 KPC 如何稳定 ADH 的可能机制。荧光猝灭数据表明，KPC 诱导动态荧光猝灭，猝灭速率常数值为 1.074 × 10 ¹⁰  M ^-1 s ^-1，表示构象变化。圆二色性进一步表明，受 KPC (5 mmol/L) 的影响，ADH 结构中可能从 α-螺旋 (14.03%–13.90%) 转变为无规卷曲 (31.98–33.04%)。分子对接分析预测KPC可能通过疏水相互作用和氢键与ADH结合。对来自 KPC 的片段肽和氨基酸的 ADH 稳定活性的验证暗示了 Cys 残基对 KPC 的生物活性的重要作用。这些结果表明KPC可能通过肽-酶相互作用稳定ADH，并保护ADH免受氧化修饰。此外，利用计算机获得了 KPC 的良好吸收、分布、代谢、排泄和毒性 (ADMET) 曲线。化验。总体而言，本研究全面了解了 KPC 和 ADH 之间的相互作用机制，并说明了 KPC 作为药物或功能性食品成分的潜在适用性。