Identification and Characterization of Lipolytic Enzymes from a Peat-Swamp Forest Soil Metagenome

• Published in: Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2010, Vol.74 (9), p.1848-1854
• Main Authors: BUNTERNGSOOK, Benjarat, KANOKRATANA, Pattanop, THONGARAM, Taksawan, TANAPONGPIPAT, Sutipa, UENGWETWANIT, Tanaporn, RACHDAWONG, Sansanalak, VICHITSOONTHONKUL, Taweerat, EURWILAICHITR, Lily
• Format: Article
• Language: English
• Subjects: acidic esterase; Biological and medical sciences; Butyrates; Cloning, Molecular; Conserved Sequence; Esterases - genetics; Esterases - isolation & purification; Esterases - metabolism; family IV lipase; Fundamental and applied biological sciences. Psychology; Industrial Microbiology; Lipase; Lipolysis; Metagenome - genetics; metagenomics; peat swamp soil; Sequence Analysis, DNA; Soil; Soil Microbiology; Substrate Specificity; Thailand; Trees; Wetlands
• ISSN: 0916-8451; 1347-6947; 1347-6947
• DOI: 10.1271/bbb.100249

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 °C at pH 5, indicating that it was an esterase with activity and secretion characteristics suitable for commercial development.