该研究以枯草芽孢杆菌和米曲霉复合发酵法制备的白芸豆多肽(white kidney bean polypeptides,WKBPs)为研究对象,利用氨基酸分析仪分析发酵前后白芸豆氨基酸组成变化,通过高效液相凝胶渗透色谱法测定WKBPs相对分子质量分布;采用体外模拟消化实验探究经胃肠道消化后WKBPs的体外降糖活性变化规律(α-淀粉酶和α-葡萄糖苷酶的抑制率),评估WKBPs在体外模拟消化系统中生物活性的稳定性。结果表明:复合发酵制备的WKBPs发酵后必需氨基酸含量从6.47 g提高到10.66 g,疏水性氨基酸含量从7.36 g增加到13.81 g,碱性氨基酸含量从2.49 g增加到3.65 g,相对分子质量分布主要集中在1 000 Da以下;α-葡萄糖苷酶和α-淀粉酶抑制率的IC50值分别为0.64和0.53 mg/mL;经胃肠模拟消化后,WKBPs对α-葡萄糖苷酶和α-淀粉酶抑制率随消化时间的增加呈先上升后下降的趋势;WKBPs的添加能够降低淀粉的体外消化性。该结果为WKBPs在营养健康食品和医药领域的进一步研究和工业应用提供科学的数据支撑。
In this study, white kidney bean polypeptides (WKBPs) were prepared by complex fermentation of Bacillus subtilis and Aspergillus oryzae.The amino acid composition and the comparative molecular weight of WKBPs were analyzed.Meanwhile, the alteration of WKBPs' hypoglycemic activity (α-glucosidase and α-amylase inhibition rate) during gastrointestinal digestion were investigated using in vitro simulated digestion system, which assessed the biological activity stability of WKBPs.The results showed that the contents of essential amino acids in WKBPs increased from 6.47 g to 10.66 g, hydrophobic amino acids increased from 7.36 g to 13.81 g, and the basic amino acids increaed from 2.49 g to 3.65 g.The relative molecular weight distribution of WKBPs was mainly below 1 000 Da.The IC50 values of WKBPs on α-glucosidase and α-amylase inhibition were 0.64 mg/mL and 0.53 mg/mL, respectively.The inhibition rate of α-glucosidase and α-amylase by WKBPs showed an increasing and then decreasing trend along with the digestion time.Meanwhile, the addition of WKBPs was able to reduce the starch digestibility in vitro.These results may provide the scientific data support for the further exploration and industrial application of WKBPs in the fields of nutrition, health food and medicine.
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