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- Publisher
- Springer Journals
- Copyright
- Copyright © 2009 by Society for Industrial Microbiology
- Subject
- Life Sciences; Microbiology ; Bioinformatics; Biotechnology; Genetic Engineering; Inorganic Chemistry ; Biochemistry, general
- ISSN
- 1367-5435
- eISSN
- 1476-5535
- DOI
- 10.1007/s10295-009-0562-7
- pmid
- 19333634
- Publisher site
- See Article on Publisher Site
Abstract
The construction of a cosmid library from the biomass produced in an enriched Sequencing Fed-Batch Reactor allowed the isolation of a new lipase by functional screening. The open reading frame of 928 bp encoded a polypeptide of 308 amino acids with a molecular mass of 32.6 kDa. The amino acid sequence analysis revealed the presence of the conserved pentapeptide GXSXG essential for lipase activity. Alignment with known sequences of proteins showed no more than 52% identity with different lipases, confirming the discovery of a novel gene sequence. The lipase was cloned and expressed in Streptomyces lividans and further purified by a combination of hydrophobic interaction and size-exclusion chromatography. Spectrophotometric assays with different p-nitrophenyl esters demonstrated a preference for long-length acyl chains, especially p-nitrophenylmyristate (C14). Moreover, the enzyme presented an optimal activity at 60°C and at alkaline pH of 10.5.
Journal
Journal of Industrial Microbiology Biotechnology – Springer Journals
Published: Mar 31, 2009