Microbial Ecology Group, Gut Health Division, Rowett Institute of Nutrition and Health, University of Aberdeen, Greenburn Road, Bucksburn, Aberdeen AB21 9SB, UK.
Environ Microbiol. 2010 Feb;12(2):304-14. doi: 10.1111/j.1462-2920.2009.02066.x. Epub 2009 Oct 5.
Butyrate-producing bacteria play an important role in the human colon, supplying energy to the gut epithelium and regulating host cell responses. In order to explore the diversity and culturability of this functional group, we designed degenerate primers to amplify butyryl-CoA:acetate CoA-transferase sequences from faecal samples provided by 10 healthy volunteers. Eighty-eight per cent of amplified sequences showed >98% DNA sequence identity to CoA-transferases from cultured butyrate-producing bacteria, and these fell into 12 operational taxonomic units (OTUs). The four most prevalent OTUs corresponded to Eubacterium rectale, Roseburia faecis, Eubacterium hallii and an unnamed cultured species SS2/1. The remaining 12% of sequences, however, belonged to 20 OTUs that are assumed to come from uncultured butyrate-producing strains. Samples taken after ingestion of inulin showed significant (P=0.019) increases in Faecalibacterium prausnitzii. Because several of the dominant butyrate producers differ in their DNA % G+C content, analysis of thermal melt curves obtained for PCR amplicons of the butyryl-CoA:acetate CoA-transferase gene provides a convenient and rapid qualitative assessment of the major butyrate producing groups present in a given sample. This type of analysis therefore provides an excellent source of information on functionally important groups within the colonic microbial community.
产丁酸菌在人类结肠中发挥着重要作用，为肠道上皮细胞提供能量，并调节宿主细胞的反应。为了探索该功能群的多样性和可培养性，我们设计了简并引物，从 10 名健康志愿者提供的粪便样本中扩增丁酰基辅酶 A：乙酸辅酶 A 转移酶序列。88%的扩增序列与培养产丁酸菌的辅酶 A 转移酶的 DNA 序列具有>98%的同一性，这些序列分为 12 个操作分类单元（OTU）。最常见的 4 个 OTU 分别对应于真杆菌属、粪真杆菌、海氏真杆菌和一个未命名的培养种 SS2/1。然而，其余 12%的序列属于 20 个假定来自未培养产丁酸菌的 OTU。摄入菊粉后采集的样本中，普雷沃氏菌属丰度显著增加（P=0.019）。由于一些主要的产丁酸菌在 DNA%G+C 含量上存在差异，因此分析丁酰基辅酶 A：乙酸辅酶 A 转移酶基因 PCR 扩增子的热融曲线可以方便、快速地定性评估给定样本中主要产丁酸菌群的存在情况。因此，这种类型的分析为结肠微生物群落中功能重要的群体提供了极好的信息来源。
