Yu-Shun Lin

Inadequate choline intake-induced choline metabolic stress (CMS) has been divergently linked to metabolic dysfunction-associated steatotic liver disease (MASLD), yet underlying mechanisms remain unclear. We hypothesized that CMS modifies plasma-free amino acid (PFAA) signatures to influence MASLD risk. In a nested case–control study of 125 participants, dietary choline intake and blood choline metabolites were assessed together with targeted metabolomics and transcriptomic profiling. MASLD was characterized by low choline intake, reduced plasma betaine/choline ratio (Pbcr), elevated homocysteine, increased branched-chain amino acids (BCAAs), and depleted serine/glycine, achieving strong predictive accuracy (AUC = 0.90). CMS was associated with reduced lymphocytic transcripts involved in BCAA catabolism and altered mTORC1/Akt/IRS1 signaling. Nonlinear Pbcr- and intake-dependent MASLD risk patterns were attenuated after adjustment for genetic–metabolite networks. These findings identify CMS-responsive metabolic mediators supporting precision choline interventions.

Dietary protein hydrolysates were evaluated as β-secretase (BACE1) inhibitors using integrated in silico, in vitro, and computational approaches. From 888 protein–enzyme combinations screened via BIOPEP-UWM, Met-containing peptide frequency (FP-M) best correlated with BACE1 inhibition (r = 0.59, p = 0.021) among 15 experimentally validated hydrolysates. LC-MS/MS confirmed 17 predicted peptide sequences from the hydrolysates. Multi-site molecular docking against four BACE1 binding sites (−8.33 to −2.88 kcal/mol) and enzyme kinetics of 10 selected peptides revealed three co-existing inhibition mechanisms: competitive (Ki = 21.2–72.0 μM, active site), mixed-type (Ki = 25.2–155.2 μM, flap/extended groove), and non-competitive (Ki = 70.4–90.8 μM, extended groove), with strong concordance between predicted binding sites and kinetic mechanisms. Molecular dynamics simulations (700 ns) confirmed mechanism-dependent binding stability; notably, the mixed-type inhibitor ENY exhibited site-dependent stability providing atomic-level evidence for dual-mode inhibition. This study establishes a validated pipeline for discovering multi-mechanism dietary BACE1 inhibitory peptides for Alzheimer's disease prevention.

Hyperuricemia is a prevalent metabolic disorder, and xanthine oxidase (XO) inhibition remains a primary strategy for its management. In this study, a novel food-derived XO inhibitory octapeptide, SRGQIEEL, was identified from yellow field pea (Pisum sativum L.) protein hydrolysates through activity-guided purification. Among six proteases tested, the Alcalase hydrolysate showed the strongest XO inhibitory activity, with an IC50 value of 0.60 mg/mL. Sequential ultrafiltration and RP-HPLC purification, followed by nanoUHPLC-ESI-Q-TOF MS, identified SRGQIEEL as the major active peptide. The synthesized peptide exhibited an IC50 of 193.34 μM and showed non-competitive inhibition kinetics, with a Ki value of 137.29 μM. Multi-site molecular docking across four cofactor-associated regions in two bovine XO structures consistently indicated that SRGQIEEL preferentially associated with the FAD domain rather than the conventional Mo-pt active site. Subsequent 100 ns molecular dynamics simulations and MM-PBSA calculations supported favorable association with the FAD region. Dynamic tunnel analysis further revealed that the FAD access tunnel remained predominantly too narrow to accommodate the peptide, supporting a tunnel-occlusion-like mechanism. Collectively, these findings suggest that SRGQIEEL inhibits XO not by entering a deeply buried active site, but by associating near the FAD-domain tunnel entrance and potentially perturbing electron transfer. This study highlights the value of dynamic tunnel accessibility analysis for investigating food-derived peptide inhibition of XO.

Advanced glycation end-products (AGEs) are undesirable compounds formed during high-temperature cooking, and charcoal-grilled meat is a major dietary source. This study developed sprayable barbecue (BBQ) sauces enriched with freeze-dried fruit powders to mitigate AGE formation in charcoal-grilled pork while maintaining product quality. Four fruit powders were initially screened and applied by marinating, spraying, or mixing. Spraying showed the most consistent inhibitory effect, and strawberry (SB) and kumquat (KQ) were selected for sauce formulation. In pork loin and belly, SB and KQ reduced Nε-carboxymethyl-lysine (CML) and Nε-carboxyethyl-lysine (CEL) by 60.9–70.5% during direct fruit powder application. Among the developed sauces, the soy sauce-free formulation showed the greatest efficacy, achieving 83.1% and 86.2% reduction in total CML and CEL with SB and KQ, respectively. All treatments significantly reduced thiobarbituric acid reactive substances (TBARS). Both formulations also maintained favorable consumer acceptance scores. Accelerated storage testing indicated that the sauces retained substantial AGE- and TBARS-inhibitory effects over 90 days at 45°C. Molecular docking suggested that 27 of 28 fruit polyphenols may bind to myosin glycation sites more favorably than AGE precursors, and methylglyoxal trapping assays indicated that B-ring hydroxylation was associated with stronger carbonyl-scavenging capacity. These findings support sprayable fruit powder BBQ sauces as a practical strategy for reducing dietary AGE exposure from charcoal-grilled pork.

Type 2 diabetes mellitus remains a critical global health challenge, driving the pursuit of novel therapeutic strategies. This study investigated the anti-diabetic efficacy of the peptide 1CBR, derived from sodium caseinate hydrolysate, administered orally at 25 mg/kg/day to db/db mice over a 4-week period. Glucose tolerance was evaluated via oral glucose tolerance tests (OGTT), while plasma dipeptidyl peptidase-IV (DPP-IV) activity, glucagon-like peptide-1 (GLP-1), and insulin concentrations were quantified using enzyme-linked immunosorbent assays (ELISA). Two bioactive peptides, GPFPLPD and APDSGNFR, were isolated and characterized, exhibiting half-maximal inhibitory concentrations (IC₅₀) of 99.12 µM and 73.07 µM for DPP-IV inhibition, respectively, and both significantly stimulated GLP-1 secretion in enteroendocrine cells in vitro. Pharmacokinetic analysis in Sprague–Dawley rats demonstrated oral bioavailability of 11.28% and 19.12% for these peptides, surpassing typical expectations for peptide-based agents. Collectively, these results provide compelling evidence that 1CBR-derived peptides exert glucose-lowering effects through the dual mechanisms of DPP-IV inhibition and GLP-1 stimulation, combined with favorable oral absorption profiles. These findings underscore the potential of 1CBR peptides as promising candidates for development into nutraceuticals or pharmaceutical agents for diabetes management.