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葡萄籽多酚纯化及体外厌氧发酵对肠道菌群的影响

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以酿酒后赤霞珠葡萄籽为原材料提取葡萄籽多酚,研究AB-8大孔吸附树脂对葡萄籽多酚的静态、动态吸附-解吸能力,筛选出适合葡萄籽多酚分离纯化的工艺条件,并在此基础上构建人粪样体外发酵模型,探讨不同质量浓度葡萄籽多酚对肠道菌群的影响.结果表明:AB-8大孔吸附树脂纯化葡萄籽多酚的最适工艺条件为:上样质量浓度8 mg/mL、乙醇体积分数80%、上样流速2mUmin、洗脱流速1 mUmin,在此纯化条件下,多酚纯度由32.16%提高至80.35%.肠道微生物测序结果显示,葡萄籽多酚质量浓度的变化并不会显著影响肠道细菌总数(P>0.05),而会显著影响肠道菌群的多样性(P<0.05).门水平下,发酵24 h后,与空白对照组(CK)相比,不同质量浓度加酚组提高了变形菌门和放线菌门的相对丰度,降低了拟杆菌门和厚壁菌门的相对丰度.低质量浓度组(LG)、中质量浓度组(MG)和高质量浓度组(HG)F/B值分别下降了8.21%,39.81%,57.49%.属水平下,多酚质量浓度的变化会影响肠道中不同菌群的生长、增殖,表现为随着质量浓度的增加对菌群有促进或抑制作用,如提高了埃希氏杆菌属/志贺氏杆菌属、克雷伯氏菌属、副杆菌属、乳酸杆菌属、双歧杆菌属、链球菌属等菌的相对丰度,降低了拟杆菌属、梭状芽胞杆菌XlVa、肠球菌属、嗜胆菌属等菌的相对丰度,随多酚质量浓度的变化表现出浓度效应,且不同发酵时间有明显波动.结论:不同质量浓度葡萄籽多酚通过调节不同菌群结构组成来改善肠道内环境,其中低质量浓度组(LG,4mg/mL)效果最好.
Effects of Purification of Grape Seed Polyphenols and in Vitro Anaerobic Fermentation on Intestinal Flora
Grape seed polyphenols were extracted from Cabernet Sauvignon grape seeds after winemaking.The static and dynamic adsorption-desorption capacity of AB-8 macroporous adsorption resin on grape seed polyphenols was studied,and the technological conditions suitable for the separation and purification of grape seed polyphenols were selected.On this basis,human fecal fermentation model was constructed in vitro to explore the effects of different concentrations of grape seed polyphenols on intestinal flora.The results showed that the optimal process conditions for purification of grape seed polyphenols by AB-8 macroporous adsorption resin were as follows:Loading mass concentration 8 mg/mL,ethanol volume fraction 80% ,loading flow rate 2 mUmin,elution flow rate 1 mUmin.Under these purification conditions,the purity of polyphenols increased from 32.16% to 80.35% .Gut microbes sequencing results showed that the grape seed polyphenols concentration changes will not significantly affect the intestinal bacteria(P>0.05),and will significantly influ-ence the diversity of intestinal flora(P<0.05).At phylum level,after 24 h of fermentation,compared with blank control group(CK),the relative abundance of Proteobacteria and Actinomycetes was increased in groups with different concentra-tions of phenol,while the relative abundance of Bacteroidetes and Firmicutes was decreased.However,F/B values in low concentration group(LG),medium concentration group(MG)and high concentration group(HG)were decreased by 8.21% ,39.81% and 57.49% ,respectively.At the genus level,the change of polyphenol concentration can affect the growth and proliferation of different intestinal flora,which could be shown as promoting or inhibiting the flora with the increase of the concentration,such as increasing the relative abundance of Escherichia/Shigella,Klebsiella,Lactobacillus,Bifidobacterium,Streptococcus and reducing the relative abundance of Bacteroides,Clostridium XlVa,Enterococcus,Bilop hila and other bacterial genera.Concentration effects were observed with changes in polyphenol mass concentration,and different fermentation time had obvious fluctuation.Therefore,different mass concentrations of grape seed polyphenols could improve the intestinal environment by regulating different bacterial community composition,and the low mass con-centration group(LG,4 mg/mL)had the best effect.

grape seed polyphenolsin vitro anaerobic fermentationintestinal florahigh-throughput sequencing

范利君、王志鹏、宋安康、李学文、王伟

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新疆农业大学食品科学与药学学院 乌鲁木齐 830000

葡萄籽多酚 体外厌氧发酵 肠道菌群 高通量测序

2024

中国食品学报
中国食品科学技术学会

中国食品学报

CSTPCD北大核心EI
影响因子:1.079
ISSN:1009-7848
年,卷(期):2024.24(8)