多菌种共酵工艺在低温长时的发酵条件下可赋予黄酒丰富的低聚糖。该研究以半干型黄酒为原料,对黄酒中低聚糖进行分离纯化及结构表征。利用75%乙醇沉淀黄酒中低聚糖,并经Sevage法除蛋白、300 Da与3 000 Da膜分离、冷冻干燥等途径获得粗黄酒低聚糖,其提取率达31.80%。粗黄酒低聚糖经琼脂糖凝胶柱DEAE Sepharose Fast Flow与葡聚糖凝胶柱Sephadex G15纯化可获得纯度为84.21%的黄酒低聚糖组分a(Huangjiu oligosaccharide-a,HJO-a)。通过建立小鼠粪便菌群体外发酵模型,探究HJO-a对肠道菌群的益生作用。结果表明,黄酒低聚糖被肠道菌群酵解后可提高发酵液中乙酸、丙酸、丁酸与异丁酸的含量,降低发酵液pH。此外,黄酒低聚糖显著提高了发酵液中肠道双歧杆菌、乳杆菌与拟杆菌的丰度(P<0.05),并显著抑制了发酵液中大肠杆菌的增殖(P<0.05)。上述结果揭示了黄酒低聚糖可发挥潜在的益生元作用。该研究为黄酒低聚糖的提取纯化及功能活性分析提供一定的理论依据。
Huangjiu is endowed with plentiful oligosaccharides in the low-temperature and long-term fermentation by a unique variety of microorganisms. In this study, Huangjiu oligosaccharides were isolated and purified from semi-dry Shaoxing Huangjiu. The oligosaccharides were extracted with 75% ethanol precipitation and Sevage method was applied to remove free proteins. Crude Huangjiu oligosaccharides were separated with 300 Da and 3000 Da membranes, subsequently freeze-drying was performed to obtain Huangjiu oligosaccharide powder, during which the extraction rate reached 31.80%. A highly-purified oligosaccharide HJO-a (purity of 84.21%) was obtained by DEAE Sepharose Fast Flow and Sephadex G15 column chromatography. An in vitro fermentation model of mouse feces was established to explore the probiotic effect of HJO-a on gut microbiota. The results showed that Huangjiu oligosaccharides were fermented by gut microbiota, which resulted in the increase of the acetic acid, propionic acid, butyric acid and isobutyric acid content, decreasing the pH of fermentation broth. Furthermore, Huangjiu oligosaccharides significantly increased the abundance of Bifidobacterium, Lactobacillus and Bacteroides (P<0.05), and evidently inhibited the proliferation of Escherichia coli in the fermentation broth (P<0.05). The aforementioned results revealed that Huangjiu oligosaccharides could exert potential prebiotic effects. The study provides a certain theoretical basis for the separation, purification, and functional activity analysis of Huangjiu oligosaccharides.
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