National Science & Technology Development Agency - Thailand; National Center Genetic Engineering & Biotechnology (BIOTEC); King Mongkuts University of Technology Thonburi
Type
Article
Source Title
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
ISSN
0916-8451
Year
2010
Volume
74
Issue
7
Open Access
Bronze
Publisher
TAYLOR & FRANCIS LTD
DOI
10.1271/bbb.100249
Format
PDF
Abstract
In this work, a metagenomic library was generated from peat-swamp forest soil obtained from Narathiwat Province, Thailand. From a fosmid library of approximately 15,000 clones, six independent clones were found to possess lipolytic activity at acidic pH. Analysis of pyrosequencing data revealed six ORFs, which exhibited 34-71% protein similarity to known lipases/esterases. A fosmid clone, designated LP8, which demonstrated the highest level of lipolytic activity under acidic conditions and demonstrated extracellular activity, was subsequently subcloned and sequenced. The full-length lipase/esterase gene, estPS2, was identified. Its deduced amino acid was closely related to a lipolytic enzyme of an uncultured bacterium, and contained the highly conserved motif of a hormone-sensitive family IV lipase. The EstPS2 enzyme exhibited highest activity toward p-nitrophenyl butyrate (C-4) at 37 degrees C at pH 5, indicating that it was an esterase with activity and secretion characteristics suitable for commercial development.
