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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Motifs
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Amino Acid Sequence
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Bacteria / genetics*
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Bacterial Proteins / chemistry
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Bacterial Proteins / genetics
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Biodiversity
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Cloning, Molecular
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Conserved Sequence
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Gene Library
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Genes, Bacterial / genetics*
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Lipolysis / genetics*
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Metagenomics / methods*
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Molecular Sequence Data
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Phylogeny
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RNA, Ribosomal, 16S / genetics
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Sewage / microbiology*
Substances
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Bacterial Proteins
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RNA, Ribosomal, 16S
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Sewage
Associated data
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GENBANK/FJ947120
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GENBANK/FJ947121
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GENBANK/FJ947122
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GENBANK/FJ947123
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GENBANK/FJ947124
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GENBANK/FJ947125
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GENBANK/FJ947126
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GENBANK/FJ947127
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GENBANK/FJ947128
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GENBANK/FJ947129
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GENBANK/FJ947130
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GENBANK/FJ947131
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GENBANK/FJ947132
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GENBANK/FJ947133
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GENBANK/FJ947134
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GENBANK/FJ947135
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GENBANK/FJ947136
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GENBANK/FJ947137
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GENBANK/FJ947138
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GENBANK/FJ947139
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GENBANK/FJ947140
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GENBANK/FJ947141
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GENBANK/FJ947142
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GENBANK/FJ947143
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GENBANK/FJ947144
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GENBANK/FJ947145
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GENBANK/FJ947146
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GENBANK/FJ947147
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GENBANK/FJ947148