title: Use of metagenomic approaches to isolate lipolytic genes from activated sludge.
abstract: 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.
authors.affiliation: Department of Agricultural Chemistry, National Taiwan University, Taipei 10617, Taiwan.
authors.forename: Ren-Bao
authors.initials: RB
authors.lastname: Liaw
authors.forename: Mei-Ping
authors.initials: MP
authors.lastname: Cheng
authors.forename: Ming-Che
authors.initials: MC
authors.lastname: Wu
authors.forename: Chia-Yin
authors.initials: CY
authors.lastname: Lee
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001419:Bacteria / Q000235:genetics*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D044822:Biodiversity; D003001:Cloning, Molecular; D017124:Conserved Sequence; D015723:Gene Library; D005798:Genes, Bacterial / Q000235:genetics*; D008066:Lipolysis / Q000235:genetics*; D056186:Metagenomics / Q000379:methods*; D008969:Molecular Sequence Data; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012722:Sewage / Q000382:microbiology*
chemical_list: D001426:Bacterial Proteins; D012336:RNA, Ribosomal, 16S; D012722:Sewage