Use of metagenomic approaches to isolate lipolytic genes from activated sludge
Use of metagenomic approaches to isolate lipolytic genes from activated sludge
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DOI：
10.1016/j.biortech.2010.05.091
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发表时间：
2010-11-01
影响因子：
11.4
通讯作者：
Lee, Chia-Yin
中科院分区：
文献类型：
--
作者：
Liaw, Ren-Bao;Cheng, Mei-Ping;Lee, Chia-Yin
本研究的目的是利用元基因组学方法从猪场废水处理设施的活性污泥中获取细菌多样性并分离脂解基因。根据对16S rRNA基因克隆的BLASTN分析，这些群落中的大多数(90%)是未培养的细菌。利用直接从活性污泥样品中提取的DNA和质粒载体构建了元基因组文库。平均插入片段大小约为5.1kb。利用三丁酸甘油酯平板法，共获得12个独特的脂解克隆。本研究中发现脂解性克隆的比率高达0.31%。分子分析显示，16个可能的脂解酶中的大多数与数据库中的非冗余蛋白质序列有28%-55%的同源性。简单地说，本研究表明活性污泥是分离新的脂解酶的理想生物资源。(C)2010爱思唯尔有限公司。保留所有权利。
The aims of this study were to access the bacterial diversity and isolate lipolytic genes using the metagenomic approach in activated sludge of a swine wastewater treatment facility. On the basis of BLASTN analysis of 16S rRNA gene clones, most of these communities (90%) were of uncultivated bacteria. The metagenomic library was constructed using a plasmid vector and DNA extracted directly from activated sludge samples. The average insert size was approximately 5.1 kb. A total of 12 unique and lipolytic clones were obtained using the tributyrin plate assay. The rate of discovering a lipolytic clone in this study was as high as 0.31%. Molecular analysis revealed that most of the 16 putative lipolytic enzymes showed 28-55% identity with non-redundant protein sequences in the database. Briefly, this study demonstrates that activated sludge is an ideal bioresource for isolating new lipolytic enzymes. (C) 2010 Elsevier Ltd. All rights reserved.