Effective lead selection for improved protein production in Aspergillus niger based on integrated genomics.

Denise I. Jacobs, Maurien M.A. Olsthoorn, Isabelle Maillet, Michiel Akeroyd, Stefaan Breestraat, Serge Donkers, Rob A.M. van der Hoeven, Cees A.M.J.J. van den Hondel, Rolf Kooistra, Thomas Lapointe, Hildegard Menke, Rogier Meulenberg, Marijke Misset, Wally H. Müller, Noël N.M.E. van Peij, Arthur Ram, Sabrina Rodriguez, Marc S. Roelofs, Johannes A. Roubos, Marcel W.E.M. van TilborgArie J. Verkleij, Herman J. Pel, Hein Stam, Cees M.J. Sagt

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Abstract

The filamentous fungus Aspergillus niger is widely exploited for industrial production of enzymes and organic acids. An integrated genomics approach was developed to determine cellular responses of A. niger to protein production in well-controlled fermentations. Different protein extraction methods in combination with automated sample processing and protein identification allowed quantitative analysis of 898 proteins. Three different enzyme overproducing strains were compared to their isogenic fungal host strains. Clear differences in response to the amount and nature of the overproduced enzymes were observed. The corresponding genes of the differentially expressed proteins were studied using transcriptomics. Genes that were up-regulated both at the proteome and transcriptome level were selected as leads for generic strain improvement. Up-regulated proteins included proteins involved in carbon and nitrogen metabolism as well as (oxidative) stress response, and proteins involved in protein folding and endoplasmic reticulum-associated degradation (ERAD). Reduction of protein degradation through the removal of the ERAD factor doaA combined with overexpression of the oligosaccharyl transferase sttC in A. niger overproducing beta-glucuronidase (GUS) strains indeed resulted in a small increase in GUS expression.

Original languageEnglish
Pages (from-to)S141-152
JournalFungal genetics and biology : FG & B
Volume46 Suppl 1
DOIs
Publication statusPublished - Mar 2009

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ASJC Scopus subject areas

  • Microbiology
  • Genetics

Cite this

Jacobs, D. I., Olsthoorn, M. M. A., Maillet, I., Akeroyd, M., Breestraat, S., Donkers, S., van der Hoeven, R. A. M., van den Hondel, C. A. M. J. J., Kooistra, R., Lapointe, T., Menke, H., Meulenberg, R., Misset, M., Müller, W. H., van Peij, N. N. M. E., Ram, A., Rodriguez, S., Roelofs, M. S., Roubos, J. A., ... Sagt, C. M. J. (2009). Effective lead selection for improved protein production in Aspergillus niger based on integrated genomics. Fungal genetics and biology : FG & B, 46 Suppl 1, S141-152. https://doi.org/10.1016/j.fgb.2008.08.012