Proteome analysis of the fungus Aspergillus carbonarius under ochratoxin A producing conditions

• Published in: International journal of food microbiology 2011-06, Vol.147 (3), p.162-169
• Main Authors: Crespo-Sempere, A., Gil, J.V., Martínez-Culebras, P.V.
• Format: Article
• Language: English
• Subjects: amino acid metabolism; amino acid sequences; Aspergillus - genetics; Aspergillus - metabolism; Aspergillus carbonarius; biosynthesis; computer software; Food Microbiology; Fungal Proteins - biosynthesis; Fungal Proteins - genetics; fungi; Gene expression; Grapes; mass spectrometry; messenger RNA; Ochratoxin production; Ochratoxins - biosynthesis; oxidative stress; Polymerase Chain Reaction; protein folding; Protein identification; protein synthesis; proteins; proteome; Proteome - genetics; Proteome - metabolism; Proteomics; RNA, Messenger - metabolism; sequence homology; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; sporulation; two-dimensional gel electrophoresis; Vitaceae; Wine; wine grapes
• ISSN: 0168-1605; 1879-3460
• DOI: 10.1016/j.ijfoodmicro.2011.03.021

Aspergillus carbonarius is an important ochratoxin A producing fungus that is responsible for mycotoxin contamination of grapes and wine. In this study, the proteomes of highly (W04-40) and weakly (W04-46) OTA-producing A. carbonarius strains were compared to identify proteins that may be involved in OTA biosynthesis. Protein samples were extracted from two biological replicates and subjected to two dimensional gel electrophoresis analysis and mass spectrometry. Expression profile comparison (PDQuest software), revealed 21 differential spots that were statistically significant and showed a two-fold change in expression, or greater. Among these, nine protein spots were identified by MALDI-MS/MS and MASCOT database and twelve remain unidentified. Of the identified proteins, seven showed a higher expression in strain W04-40 (high OTA producer) and two in strain W04-46 (low OTA producer). Some of the identified amino acid sequences shared homology with proteins involved in regulation, amino acid metabolism, oxidative stress and sporulation. It is worth noting the presence of a protein with 126.5 fold higher abundance in strain W04-40 showing homology with protein CipC, a protein with unknown function related with pathogenesis and mycotoxin production by some authors. Variations in protein expression were also further investigated at the mRNA level by real-time PCR analysis. The mRNA expression levels from three identified proteins including CipC showed correlation with protein expression levels. This study represents the first proteomic analysis for a comparison of two A. carbonarius strains with different OTA production and will contribute to a better understanding of the molecular events involved in OTA biosynthesis. ►The proteomes of highly and weakly OTA-producing A. carbonarius strains were compared. ►Nine differential protein spots were identified by MALDI-MS/MS and MASCOT. ►Identified proteins were involved in regulation, amino acid metabolism, oxidative stress and sporulation. ►A protein with 126.5 fold higher abundance in high OTA-producing strain showed homology with CipC. ►mRNA variations were also investigated by real-time PCR analysis.