Tricholoma matsutake Protein as a Novel Cognitive Supportive Nutrient: Identification and Molecular Interaction...

Maintaining or enhancing cognitive function in healthy individuals is essential for preventing neurodegenerative diseases, and increasing evidence highlights the link between dietary supplementation and cognitive health. Recent studies have mainly focused on alleviating mild cognitive impairment (MCI) through edible mushroom supplementation, yet substantial knowledge gaps remain. Therefore, this study examined the effects of daily dietary supplementation with Tricholoma matsutake powder (TP) and its major bioactive components─proteins (TPT) and polysaccharides (TPS)─on mouse cognitive function and identified novel memory-enhancing peptides within TPT, which showed the strongest efficacy. After 30 days of supplementation, mice receiving TP, TPT, or TPS showed superior performance in object recognition, spatial learning, and working memory compared with controls, with TPT demonstrating the most pronounced improvement. TPT better promoted the morphological integrity of hippocampal neurons, increased Nissl body density, enhanced cholinergic function, and markedly reduced oxidative stress. Furthermore, liquid chromatography-tandem mass spectrometry analysis identified 2287 peptides in TPT digestion products. In vitro digestion and absorption assays further screened 168 stable peptides with potential memory-enhancing activity. Molecular docking revealed multiple interactions between Pro-Phe-Leu (PFL), Leu- Phe-Leu (LFL), Ala- Phe-Leu (AFL), and acetylcholinesterase (AChE) binding sites, with high docking scores and stable binding, suggesting that these peptides may serve as potent AChE inhibitors. These findings indicate that dietary TPT supplementation enhances memory function in mice, possibly mediated by bioavailable active peptides. PFL, LFL, and AFL may be more promising candidate molecules for preventing MCI.