Identification and characterization of lipolytic enzymes from a peat-swamp forest soil metagenome.
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.
Bunterngsook
Benjarat
B
Division of Biotechnology, School of Bioresources and Technology, King Mongkut's University of Technology Thonburi, Bangkok, Thailand.
Kanokratana
Pattanop
P
Thongaram
Taksawan
T
Tanapongpipat
Sutipa
S
Uengwetwanit
Tanaporn
T
Rachdawong
Sansanalak
S
Vichitsoonthonkul
Taweerat
T
Eurwilaichitr
Lily
L
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
D002087:Butyrates; D012987:Soil; C033592:4-nitrophenyl butyrate; D004950:Esterases; D008049:Lipase