Journal
BIORESOURCE TECHNOLOGY
Volume 101, Issue 21, Pages 8323-8329Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.biortech.2010.05.091
Keywords
Lipolytic genes; Activated sludge; Metagenome
Funding
- National Science Council [NSC98-2313-B-002-052-MY3]
- Council of Agriculture, Taiwan [COA98AS-8.1.3-LI-L1]
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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.
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