Novel hydrolase diversity retrieved from a metagenome library of bovine rumen microflora
Novel hydrolase diversity retrieved from a metagenome library of bovine rumen microflora
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DOI：
10.1111/j.1462-2920.2005.00920.x
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发表时间：
2005-12-01
影响因子：
5.1
通讯作者：
Golyshin, PN
中科院分区：
文献类型：
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作者：
Ferrer, M;Golyshina, OV;Golyshin, PN
在噬菌体 lambda 载体中建立了从奶牛瘤胃内容物中提取的大量 DNA 的宏基因组表达文库，并采用基于活性的筛选来探索微生物菌群的功能多样性。在文库中鉴定并鉴定了 22 个具有不同水解活性的克隆（12 种酯酶、9 种内切 β-1,4-葡聚糖酶和 1 种环糊精酶）。对检索到的酶的序列分析显示，其中八种（36%）是全新的，并形成了深分支的系统发育谱系，在已知的酯水解酶和糖基水解酶中没有近亲。对水解酶基因序列以及邻近基因的序列和背景的生物信息学分析表明，对产生回收的酶的瘤胃生物体进行了初步的系统发育分配。水解酶的系统发育新颖性表明其中一些酶可能具有生物催化新应用的潜力。
A metagenome expression library of bulk DNA extracted from the rumen content of a dairy cow was established in a phage lambda vector and activity-based screening employed to explore the functional diversity of the microbial flora. Twenty-two clones specifying distinct hydrolytic activities ( 12 esterases, nine endo-beta-1,4-glucanases and one cyclodextrinase) were identified in the library and characterized. Sequence analysis of the retrieved enzymes revealed that eight (36%) were entirely new and formed deep-branched phylogenetic lineages with no close relatives among known ester- and glycosyl-hydrolases. Bioinformatic analyses of the hydrolase gene sequences, and the sequences and contexts of neighbouring genes, suggested tentative phylogenetic assignments of the rumen organisms producing the retrieved enzymes. The phylogenetic novelty of the hydrolases suggests that some of them may have potential for new applications in biocatalysis.