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采用 16S rRNA 基因克隆文库揭示了传统中国后发酵普洱茶发酵过程中细菌群落的结构和动态。

Structure and dynamics of the bacterial communities in fermentation of the traditional Chinese post-fermented pu-erh tea revealed by 16S rRNA gene clone library.

机构信息

College of Longrun Pu-erh Tea, Yunnan Agricultural University, Kunming, 650201, Yunnan, China,

出版信息

World J Microbiol Biotechnol. 2013 Oct;29(10):1877-84. doi: 10.1007/s11274-013-1351-z. Epub 2013 Apr 17.

Abstract

Microbes are thought to have key roles in the development of the special properties of post-fermented pu-erh tea (pu-erh shucha), a well-known traditional Chinese tea; however, little is known about the bacteria during the fermentation. In this work, the structure and dynamics of the bacterial community involved in the production of pu-erh shucha were investigated using 16S rRNA gene clone libraries constructed from samples collected on days zero (LD-0), 5 (LD-5), 10 (LD-10), 15 (LD-15) and 20 (LD-20) of the fermentation. A total of 747 sequences with individual clone library containing 115-174 sequences and 4-20 unique operational taxonomic units (OTUs) were obtained. These OTUs were grouped into four phyla (Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria) and further identified as members of 10 families, such as Alcaligenaceae, Bacillaceae, Enterobacteriaceae, etc. The dominant bacteria were Enterobacteriaceae in the raw material (LD-0) and in the initial stages of fermentation (LD-5 and LD-10), which changed to Bacillaceae at the last stages of fermentation (LD-15 and LD-20) at a temperature of 40-60 °C. It is interesting that the dominant OTUs in libraries LD-15 and LD-20 were very closely related to Bacillus coagulans, which is a safe thermoduric probiotic. Together the bacterial diversity and dynamics during a fermentation of pu-erh shucha were demonstrated, and a worthy clue for artificial inoculation of B. coagulans to improve the health benefits of pu-erh shucha or produce probiotic pu-erh tea were provided.

摘要

微生物被认为在后期发酵普洱茶(普洱熟茶)特殊性质的形成中起着关键作用,这是一种著名的中国传统茶;然而,对于发酵过程中的细菌知之甚少。在这项工作中,使用从发酵第 0 天(LD-0)、第 5 天(LD-5)、第 10 天(LD-10)、第 15 天(LD-15)和第 20 天(LD-20)收集的样品构建的 16S rRNA 基因克隆文库,研究了参与普洱熟茶生产的细菌群落的结构和动态。总共获得了 747 个序列,每个克隆文库包含 115-174 个序列和 4-20 个独特的操作分类单元(OTU)。这些 OTU 被分为 4 个门(放线菌门、拟杆菌门、厚壁菌门和变形菌门),并进一步鉴定为 10 个科的成员,如产碱杆菌科、芽孢杆菌科、肠杆菌科等。在原料(LD-0)和发酵初期(LD-5 和 LD-10),优势细菌是肠杆菌科,在发酵后期(LD-15 和 LD-20)温度为 40-60°C 时变为芽孢杆菌科。有趣的是,LD-15 和 LD-20 文库中的优势 OTU 与凝结芽孢杆菌非常密切相关,凝结芽孢杆菌是一种安全的耐热益生菌。该研究展示了普洱熟茶发酵过程中的细菌多样性和动态,为人工接种凝结芽孢杆菌以提高普洱熟茶的健康益处或生产益生菌普洱茶提供了一个有价值的线索。

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