Yang Shuanghong, Xie Xinqiang, Ma Jun, He Xingxiang, Li Ying, Du Mingzhu, Li Longyan, Yang Lingshuang, Wu Qingping, Chen Wei, Zhang Jumei
School of Food Science and Technology, Jiangnan University, Wuxi, China.
Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou, China.
Front Microbiol. 2021 Apr 21;12:649698. doi: 10.3389/fmicb.2021.649698. eCollection 2021.
, an important genus for human health, is difficult to isolate. We applied metagenomics, pangenomics, and enzymology to determine the dominant glycoside hydrolase (GH) families of Bifidobacterium and designed selective medium for isolation. Pangenomics results showed that the GH13, GH3, GH42, and GH43 families were highly conserved in . Metagenomic analysis of GH families in human faecal samples was performed. The results indicated that contains core GHs for utilizing raffinose, D-trehalose anhydrous, D(+)-cellobiose, melibiose, lactulose, lactose, D(+)-sucrose, resistant starch, pullulan, xylan, and glucan. These carbohydrates as the main carbon sources were applied for selective media, which were more conducive to the growth of bifidobacteria. In the medium with lactose, raffinose and xylan as the main carbon sources, the ratio of cultivable bifidobacteria to cultivable microorganisms were 89.39% ± 2.50%, 71.45% ± 0.99%, and 53.95% ± 1.22%, respectively, whereas the ratio in the ordinary Gifu anaerobic medium was only 17.90% ± 0.58%. Furthermore, the species significantly ( < 0.05) varied among samples from different individuals. Results suggested that xylan might be a prebiotic that benefits host health, and it is feasible to screen and isolate bifidobacteria using the oligosaccharides corresponding to the specific GHs of bifidobacteria as the carbon sources of the selective media.
双歧杆菌属是对人类健康很重要的一个属,但难以分离。我们应用宏基因组学、泛基因组学和酶学来确定双歧杆菌的主要糖苷水解酶(GH)家族,并设计了用于分离的选择性培养基。泛基因组学结果表明,GH13、GH3、GH42和GH43家族在双歧杆菌属中高度保守。对人类粪便样本中的GH家族进行了宏基因组分析。结果表明,双歧杆菌属含有用于利用棉子糖、无水D -海藻糖、D(+) -纤维二糖、蜜二糖、乳果糖、乳糖、D(+) -蔗糖、抗性淀粉、支链淀粉、木聚糖和葡聚糖的核心GHs。这些碳水化合物作为主要碳源被应用于选择性培养基,这更有利于双歧杆菌的生长。在以乳糖、棉子糖和木聚糖作为主要碳源的培养基中,可培养双歧杆菌与可培养微生物的比例分别为89.39% ± 2.50%、71.45% ± 0.99%和53.95% ± 1.22%,而在普通岐阜厌氧培养基中的比例仅为17.90% ± 0.58%。此外,不同个体样本中的物种存在显著差异(P < 0.05)。结果表明木聚糖可能是一种有益于宿主健康的益生元,使用与双歧杆菌特定GHs相对应的低聚糖作为选择性培养基的碳源来筛选和分离双歧杆菌是可行的。