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Current Genetics

, Volume 59, Issue 3, pp 129–137 | Cite as

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

  • Suja MohammedEmail author
  • Junior Te’o
  • Helena Nevalainen
Research Article

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.

Keywords

Cold-active lipase Antarctic Penicillium expansum Genomic walking PCR 

Notes

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.

Supplementary material

294_2013_394_MOESM1_ESM.doc (841 kb)
Supplementary material 1 (DOC 841 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Suja Mohammed
    • 1
    Email author
  • Junior Te’o
    • 2
  • Helena Nevalainen
    • 2
  1. 1.Australian School of Advanced Medicine, Faculty of Human SciencesMacquarie UniversitySydneyAustralia
  2. 2.Department of Chemistry and Biomolecular Sciences, Biomolecular Frontiers Research CentreMacquarie UniversitySydneyAustralia

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