title: Identification and characterization of lipolytic enzymes from a peat-swamp forest soil metagenome.
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 (C⁴) at 37 °C at pH 5, indicating that it was an esterase with activity and secretion characteristics suitable for commercial development.
authors.affiliation: Division of Biotechnology, School of Bioresources and Technology, King Mongkut's University of Technology Thonburi, Bangkok, Thailand.
authors.forename: Benjarat
authors.initials: B
authors.lastname: Bunterngsook
authors.forename: Pattanop
authors.initials: P
authors.lastname: Kanokratana
authors.forename: Taksawan
authors.initials: T
authors.lastname: Thongaram
authors.forename: Sutipa
authors.initials: S
authors.lastname: Tanapongpipat
authors.forename: Tanaporn
authors.initials: T
authors.lastname: Uengwetwanit
authors.forename: Sansanalak
authors.initials: S
authors.lastname: Rachdawong
authors.forename: Taweerat
authors.initials: T
authors.lastname: Vichitsoonthonkul
authors.forename: Lily
authors.initials: L
authors.lastname: Eurwilaichitr
mesh_terms: D002087:Butyrates; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004950:Esterases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D007218:Industrial Microbiology; D008049:Lipase; D008066:Lipolysis; D054892:Metagenome / Q000235:genetics*; D017422:Sequence Analysis, DNA; D012987:Soil; D012988:Soil Microbiology; D013379:Substrate Specificity; D013785:Thailand; D014197:Trees; D053833:Wetlands
chemical_list: D002087:Butyrates; D012987:Soil; C033592:4-nitrophenyl butyrate; D004950:Esterases; D008049:Lipase