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A gene encoding a new cold-active lipase from an Antarctic isolate of Penicillium expansum

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Abstract

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.

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Acknowledgments

We thank Paul Worden from the Macquarie University Sequencing Facility for DNA sequencing. We also thank Dr. Ron Bradner for sharing information about the Antarctic fungal strains and identification of strains. S. Mohammed would like to express her sincere gratitude to Prof. Paul Pilowsky for his enormous support and valuable advice on manuscript preparation. This research has been facilitated by access to the Australian Proteome Analysis Facility established under the Australian Government’s Major National Research Facilities Program. S. Mohammed is the recipient of a RAACE (Research Award in Areas and Centres of Excellence) Scholarship awarded by Macquarie University.

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Authors and Affiliations

  1. Australian School of Advanced Medicine, Faculty of Human Sciences, Macquarie University, Sydney, NSW, 2109, Australia

    Suja Mohammed

  2. Department of Chemistry and Biomolecular Sciences, Biomolecular Frontiers Research Centre, Macquarie University, Sydney, NSW, 2109, Australia

    Junior Te’o & Helena Nevalainen

Authors
  1. Suja Mohammed
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  2. Junior Te’o
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  3. Helena Nevalainen
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Corresponding author

Correspondence to Suja Mohammed.

Additional information

Communicated by A. Brakhage.

Nucleotide sequence data reported in this study are available in the GenBank database under the accession number KC195873.

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Mohammed, S., Te’o, J. & Nevalainen, H. A gene encoding a new cold-active lipase from an Antarctic isolate of Penicillium expansum . Curr Genet 59, 129–137 (2013). https://doi.org/10.1007/s00294-013-0394-x

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  • DOI: https://doi.org/10.1007/s00294-013-0394-x

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