Identification and characterization of lipolytic enzymes from a peat-swamp forest soil metagenome
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Author list: Bunterngsook B., Kanokratana P., Thongaram T., Tanapongpipat S., Uengwetwanit T., Rachdawong S., Vichitsoonthonkul T., Eurwilaichitr L.
Publisher: Taylor & Francis: STM, Behavioural Science and Public Health Titles
Publication year: 2010
Journal: Bioscience, Biotechnology and Biochemistry (0916-8451)
Volume number: 74
Issue number: 9
Start page: 1848
End page: 1854
Number of pages: 7
ISSN: 0916-8451
Languages: English-Great Britain (EN-GB)
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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 Upases/ 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 /7-nitrophenyl butyrate (C4) at 37 ฐC at pH 5, indicating that it was an esterase with activity and secretion characteristics suitable for commercial development.
Keywords
Acidic esterase, Family IV lipase, Peat swamp soil
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