滇金丝猴粪便微生物GH10家族木聚糖酶基因多样性

Gene diversity of the glycosyl hydrolase family 10 xylanase in the fecal microorganism of Rhinopithecus bieti

摘要:

【目的】分析滇金丝猴粪便微生物中GH10家族木聚糖酶的基因多样性。【方法】以野生和半圈养滇金丝猴粪便微生物宏基因组DNA为模板，用GH10木聚糖酶简并引物扩增木聚糖酶基因片段，利用pMD19-T载体构建细菌克隆文库并进行分析。【结果】从野生和半圈养滇金丝猴粪便微生物克隆文库中分别获得26、28条GH10木聚糖酶基因片段，与GenBank中木聚糖酶序列一致性分别介于58%?95%、63%?81%之间。比对分析表明，两种环境中的GH10木聚糖酶均来自厚壁菌门、拟杆菌门和未培养细菌。野生滇金丝猴粪便微生物的GH10木聚糖酶基因来源于uncultured bacterium和Butyrivibrio、Bacteroides、Ruminococcus、Sphingobacterium、Chryseobacterium、Clostridium、Bacillus 7个属；而半圈养滇金丝猴粪便微生物的GH10木聚糖酶基因来源于uncultured bacterium和Clostridium、Paludibacter、Sphingobacterium、Ruminococcus、Roseburia、Chryseobacterium 6个属，其中两种环境都存在来源于Ruminococcus、Clostridium、Chryseobacterium、Sphingobacterium的GH10木聚糖酶。【结论】滇金丝猴粪便微生物中含有丰富的GH10木聚糖酶基因，且野生和半圈养两种不同环境中GH10木聚糖酶基因的微生物来源存在一定差异。该研究丰富了动物胃肠道中GH10木聚糖酶基因多样性，并为新型木聚糖酶的开发和滇金丝猴胃肠道微生物资源的利用奠定了基础。

Abstract:

[Objective] The diversity of the glycosyl hydrolase family 10 (GH10) xylanase gene of fecal microorganism of Rhinopithecus bieti is studied. [Methods] Using the genomic DNA extracted from the feces of wild and half-wild R. bieti as template, we amplified xylanase gene fragments by degenerate primer of GH10 xylanase. Bacteria clone library is constructed and analyzed using pMD19-T vector. [Results] From fecal microbial clone library of wild and half-wild R. bieti 26 and 28 GH10 xylanase gene fragments are respectively acquired, whose identity with xylanase sequences in GenBank varies from 58%?95% to 63%?81%. Blast results shows that microbial composition of GH10 xylanase from two different environments is similar, including the Firmicutes, Bacteroidetes and uncultured bacteria. GH10 xylanase gene of fecal microbiome of wild R. bieti derives from Uncultured bacterium, Butyrivibrio, Bacteroides, Ruminococcus, Sphingobacterium, Chryseobacterium, Clostridium, Bacillus, while that of half-wild R. bieti only from Uncultured bacterium, Clostridium, Paludibacter, Sphingobacterium, Ruminococcus, Roseburia, Chryseobacterium. There is GH10 xylanase existing in both environments, including Ruminococcus, Clostridium, Chryseobacterium, Sphingobacterium. [Conclusion] There are abundant GH10 xylanase genes in fecal microorganism of wild and half-wild R. bieti and there is some difference in their microbial sources. This research enriches the diversity of GH10 xylanase in animal gastrointestinal tract as well as sets foundations for the exploitation of novel xylanase and the utilization of microbial resources in R. bieti’s gastrointestinal tract.