A gene encoding a new cold-active lipase from an Antarctic isolate of Penicillium expansum
| Autor: | Suja Mohammed, Junior Te’o, Helena Nevalainen |
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| Rok vydání: | 2013 |
| Předmět: |
Signal peptide
DNA Complementary Antarctic Regions Tandem Mass Spectrometry Complementary DNA Enzyme Stability Genetics Escherichia coli Amino Acid Sequence Lipase Cloning Molecular Peptide sequence Gene chemistry.chemical_classification biology Sequence Homology Amino Acid Penicillium Temperature food and beverages General Medicine biology.organism_classification Molecular biology Amino acid genomic DNA Biochemistry chemistry biology.protein Penicillium expansum |
| Zdroj: | Current genetics. 59(3) |
| ISSN: | 1432-0983 |
| Popis: | Cold-active lipases are of significant interest as biocatalysts in industrial processes. We have identified a lipase that displayed activity towards long carbon-chain-p-nitrophenyl substrates (C12–C18) at 25 °C from the culture supernatant of an Antarctic Penicillium expansum strain assigned P. expansum SM3. Zymography revealed a protein band of around 30 kDa with activity towards olive oil. DNA fragments of a lipase gene designated as lipPE were isolated from the genomic DNA of P. expansum SM3 by genomic walking PCR. Subsequently, the complete genomic lipPE gene was amplified using gene-specific primers designed from the 5′- and 3′-regions. Reverse transcription PCR was used to amplify the lipPE cDNA. The deduced amino acid sequence consisted of 285 residues that included a predicted signal peptide. Three peptides identified by LC/MS/MS analysis of the proteins in the culture supernatant of P. expansum were also present in the deduced amino acid sequence of the lipPE gene suggesting that this gene encoded the lipase identified by initial zymogram activity analysis. Full analysis of the nucleotide and the deduced amino acid sequences indicated that the lipPE gene encodes a novel P. expansum lipase. The lipPE gene was expressed in E. coli for further characterization of the enzyme with a view of assessing its suitability for industrial applications. |
| Databáze: | OpenAIRE |
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