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

被引：47

作者：

Jacobs, Denise I. ^{[1
]}

Olsthoorn, Maurien M. A. ^{[1
]}

Maillet, Isabelle ^{[2
]}

Akeroyd, Michiel ^{[1
]}

Breestraat, Stefaan ^{[1
]}

Donkers, Serge ^{[1
]}

van der Hoeven, Rob A. M. ^{[1
]}

van den Hondel, Cees A. M. J. J. ^{[3
]}

Kooistra, Rolf ^{[3
]}

Lapointe, Thomas ^{[2
]}

Menke, Hildegard ^{[1
]}

Meulenberg, Rogier ^{[1
]}

Misset, Marijke ^{[1
]}

Mueller, Wally H. ^{[4
]}

van Peij, Noel N. M. E. ^{[1
]}

Ram, Arthur ^{[3
]}

Rodriguez, Sabrina ^{[2
]}

Roelofs, Marc S. ^{[4
]}

Roubos, Johannes A. ^{[1
]}

van Tilborg, Marcel W. E. M. ^{[1
]}

Verkleij, Arie J. ^{[4
]}

Pel, Herman J. ^{[1
]}

Stam, Hein ^{[1
]}

Sagt, Cees M. J. ^{[1
]}

机构：

[1] DSM Food Specialties, NL-2600 MA Delft, Netherlands

[2] DSM Nutr Prod, CH-4002 Basel, Switzerland

[3] Leiden Univ, NL-2333 AL Leiden, Netherlands

[4] Univ Utrecht, NL-3584 CH Utrecht, Netherlands

来源：

FUNGAL GENETICS AND BIOLOGY | 2009年 / 46卷

关键词：

Aspergillus niger; Proteomics; Transcriptomics; Protein production; Secretion stress; Oxidative stress; Fermentation; sttC; doaA; ERAD; ENDOPLASMIC-RETICULUM ER; SACCHAROMYCES-CEREVISIAE; FILAMENTOUS FUNGI; 2-DIMENSIONAL ELECTROPHORESIS; ELECTRON MICROSCOPY; OXIDATIVE STRESS; CELL-WALL; PROTEOMICS; SECRETION; NIDULANS;

D O I：

10.1016/j.fgb.2008.08.012

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

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 over-producing beta-glucuronidase (GUS) strains indeed resulted in a small increase in GUS expression. (C) 2008 Elsevier Inc. All rights reserved.

引用

页码：S141 / S152

页数：12

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