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Mycotoxin Research

, Volume 26, Issue 3, pp 171–180 | Cite as

Proteome analysis of Aspergillus ochraceus

  • Muhammad Rizwan
  • Ingrid Miller
  • Fareeha Tasneem
  • Josef Böhm
  • Manfred Gemeiner
  • Ebrahim Razzazi-FazeliEmail author
Original Paper

Abstract

Genome sequencing for many important fungi has begun during recent years; however, there is still some deficiency in proteome profiling of aspergilli. To obtain a comprehensive overview of proteins and their expression, a proteomic approach based on 2D gel electrophoresis and MALDI-TOF/TOF mass spectrometry was used to investigate A. ochraceus. The cell walls of fungi are exceptionally resistant to destruction, therefore two lysis protocols were tested: (1) lysis via manual grinding using liquid nitrogen, and (2) mechanical lysis via rapid agitation with glass beads using MagNalyser. Mechanical grinding with mortar and pestle using liquid nitrogen was found to be a more efficient extraction method for our purpose, resulting in extracts with higher protein content and a clear band pattern in SDS-PAGE. Two-dimensional electrophoresis gave a complex spot pattern comprising proteins of a broad range of isoelectric points and molecular masses. The most abundant spots were subjected to mass spectrometric analysis. We could identify 31 spots representing 26 proteins, most of them involved in metabolic processes and response to stress. Seventeen spots were identified by de novo sequencing due to a lack of DNA and protein database sequences of A. ochraceus. The proteins identified in our study have been reported for the first time in A. ochraceus and this represents the first proteomic approach with identification of major proteins, when the fungus was grown under submerged culture.

Keywords

Proteome Aspergillus ochraceus Two-dimensional Gel electrophoresis MALDI-TOF/TOF mass spectrometer Ochratoxin A De novo sequencing 

Notes

Acknowledgements

The authors are grateful to the Higher Education Commission (HEC) Islamabad, Pakistan and Austrian Exchange Service (ÖAD) Vienna, Austria for providing a scholarship to Mr. Muhammad Rizwan.

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Copyright information

© Society for Mycotoxin Research and Springer 2010

Authors and Affiliations

  • Muhammad Rizwan
    • 1
  • Ingrid Miller
    • 2
  • Fareeha Tasneem
    • 1
  • Josef Böhm
    • 3
  • Manfred Gemeiner
    • 2
  • Ebrahim Razzazi-Fazeli
    • 1
    • 3
    Email author
  1. 1.VetOMICS Core Facility for ResearchUniversity of Veterinary Medicine ViennaViennaAustria
  2. 2.Department for Biomedical SciencesUniversity of Veterinary Medicine ViennaViennaAustria
  3. 3.Department of Public Health in Veterinary MedicineUniversity of Veterinary Medicine ViennaViennaAustria

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