Ferrer Manuel, Golyshina Olga V, Chernikova Tatyana N, Khachane Amit N, Reyes-Duarte Dolores, Santos Vitor A P Martins Dos, Strompl Carsten, Elborough Kieran, Jarvis Graeme, Neef Alexander, Yakimov Michail M, Timmis Kenneth N, Golyshin Peter N
Department of Microbiology, GBF- German Research Centre for Biotechnology, Mascheroder Weg 1, 38124 Braunschweig, Germany.
Environ Microbiol. 2005 Dec;7(12):1996-2010. doi: 10.1111/j.1462-2920.2005.00920.x.
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.
利用噬菌体λ载体构建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方法来探索微生物群落的功能多样性。在该文库中鉴定并表征了22个具有不同水解活性的克隆（12个酯酶、9个内切β-1,4-葡聚糖酶和1个环糊精酶）。对所获得酶的序列分析表明，其中8个（36%）是全新的，在已知的酯酶和糖基水解酶中没有近亲，形成了深分支的系统发育谱系。对水解酶基因序列以及相邻基因的序列和背景进行生物信息学分析，对产生所获得酶的瘤胃微生物进行了初步的系统发育定位。水解酶在系统发育上的新颖性表明，其中一些可能在生物催化中有新的应用潜力。
