Identification and characterization of lipolytic enzymes from a peat-swamp forest soil metagenome
| Autor: | Benjarat Bunterngsook, Pattanop Kanokratana, Lily Eurwilaichitr, Taksawan Thongaram, Sutipa Tanapongpipat, Taweerat Vichitsoonthonkul, Sansanalak Rachdawong, Tanaporn Uengwetwanit |
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| Rok vydání: | 2010 |
| Předmět: |
Esterase Gene
Lipolysis Triacylglycerol lipase Applied Microbiology and Biotechnology Biochemistry Esterase Analytical Chemistry Microbiology Substrate Specificity Trees Industrial Microbiology Soil Lipase Cloning Molecular Molecular Biology Conserved Sequence Soil Microbiology biology Organic Chemistry Esterases General Medicine Sequence Analysis DNA biology.organism_classification Thailand Enzyme assay Fosmid Butyrates Wetlands biology.protein Metagenome Soil microbiology Bacteria Biotechnology |
| Zdroj: | Bioscience, biotechnology, and biochemistry. 74(9) |
| ISSN: | 1347-6947 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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