Bile acid (BA) micelle-disrupting peptides are bioactive peptides (BPs) that suppress intestinal cholesterol adsorption and ameliorate blood cholesterol levels. To facilitate their application in the food industry, an effective and straightforward technology for BP separation and enrichment of BPs is required. For this purpose, we assessed the potential of heat-treated (HT) porous silica gels with an enhanced capacity for peptide adsorption/desorption (AD). Given the key characteristics of these peptides involving basic amino acid (AA) residues, we applied casein hydrolysate to HT silica gels for adsorption at neutral pH, where the surface yielded anionic silanolates through deprotonation. Subsequently, desorption was conducted at pH 2, at which point the surface returned to its neutral state. We confirmed a 335-fold increase in the BA micelle-disrupting activity of the resulting hydrolysates. LC-MS/MS analysis identified 783 peptides, confirming that the peptides containing cationic AA residues were selectively enriched, as expected. Moreover, a dose-response of randomly selected 27 peptides with various enrichment levels revealed the presence of multiple active peptides in the enriched groups. These results indicate that our method is effective for the separation and enrichment of BA micelle-disrupting peptides and holds promise for BP production.

中文翻译：

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利用热处理多孔硅胶吸附/解吸过程分离和富集多种胆汁酸胶束破坏肽

胆汁酸 (BA) 胶束破坏肽是生物活性肽 (BP)，可抑制肠道胆固醇吸附并改善血液胆固醇水平。为了促进其在食品工业中的应用，需要一种有效且简单的 BP 分离和浓缩技术。为此，我们评估了热处理 (HT) 多孔硅胶的潜力，该硅胶具有增强的肽吸附/解吸 (AD) 能力。鉴于这些肽涉及碱性氨基酸 (AA) 残基的关键特征，我们将酪蛋白水解物应用到 HT 硅胶上，在中性 pH 值下进行吸附，其中表面通过去质子化产生阴离子硅烷醇盐。随后，在pH 2下进行解吸，此时表面恢复到中性状态。我们证实所得水解产物的 BA 胶束破坏活性增加了 335 倍。 LC-MS/MS 分析鉴定出 783 种肽，证实含有阳离子 AA 残基的肽如预期那样被选择性富集。此外，随机选择的 27 种具有不同富集水平的肽的剂量反应表明，富集组中存在多种活性肽。这些结果表明我们的方法可有效分离和富集 BA 胶束破坏肽，并有望用于 BP 生产。