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来自突尼斯单峰驼粪便微生物群的两个参与植物细胞壁降解的新基因簇。

Two new gene clusters involved in the degradation of plant cell wall from the fecal microbiota of Tunisian dromedary.

机构信息

Laboratoire de Biotechnologie Microbienne et d'Ingénierie des Enzymes (LBMIE), Centre of Biotechnology of Sfax (CBS), University of Sfax, Sfax, Tunisia.

LISBP, CNRS, INRA, INSAT, Université de Toulouse, Toulouse, France.

出版信息

PLoS One. 2018 Mar 30;13(3):e0194621. doi: 10.1371/journal.pone.0194621. eCollection 2018.

Abstract

Dromedaries are capable of digesting plant cell wall with high content of lignocellulose of poor digestibility. Consequently, their intestinal microbiota can be a source of novel carbohydrate-active enzymes (CAZymes). To the best of our knowledge, no data are available describing the biochemical analysis of enzymes in dromedary intestinal microbiota. To investigate new hydrolytic enzymes from the dromedary gut, a fosmid library was constructed using metagenomic DNA from feces of non-domestic adult dromedary camels living in the Tunisian desert. High-throughput functional screening of 13756 clones resulted in 47 hit clones active on a panel of various chromogenic and non-chromogenic glycan substrates. Two of them, harboring multiple activities, were retained for further analysis. Clone 26H3 displayed activity on AZO-CM-cellulose, AZCL Carob galactomannan and Tween 20, while clone 36A23 was active on AZCL carob galactomannan and AZCL barley β-glucan. The functional annotation of their sequences highlighted original metagenomic loci originating from bacteria of the Bacteroidetes/Chlorobi group, involved in the metabolization of mannosides and β-glucans thanks to a complete battery of endo- and exo-acting glycoside hydrolases, esterases, phosphorylases and transporters.

摘要

骆驼能够消化富含木质纤维素、消化率低的植物细胞壁。因此,它们的肠道微生物群可能是新型碳水化合物活性酶(CAZymes)的来源。据我们所知,目前尚无描述骆驼肠道微生物群中酶的生化分析的数据。为了从骆驼肠道中研究新的水解酶,我们使用来自生活在突尼斯沙漠中的非家养成年单峰驼粪便中的宏基因组 DNA 构建了一个 fosmid 文库。对 13756 个克隆进行高通量功能筛选,结果得到了 47 个在各种显色和非显色糖基底物上具有活性的克隆。其中两个具有多种活性的克隆被保留用于进一步分析。克隆 26H3 在 AZO-CM-纤维素、AZCL 角豆半乳甘露聚糖和吐温 20 上具有活性,而克隆 36A23 在 AZCL 角豆半乳甘露聚糖和 AZCL 大麦β-葡聚糖上具有活性。它们序列的功能注释突出了来源于拟杆菌门/绿菌门细菌的原始宏基因组基因座,这些基因座通过一系列内、外作用糖苷水解酶、酯酶、磷酸酶和转运体参与甘露糖苷和β-葡聚糖的代谢。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a47c/5877837/3b329a5e358c/pone.0194621.g001.jpg

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