The synergetic effects of two CCAAT boxes in Aspergillus niger glaA gene promoter on activation of PglaA transcription

2004 ◽  
Vol 47 (2) ◽  
pp. 139 ◽  
Author(s):  
Xingguo ZHU
Keyword(s):  
Aspergillus Niger ◽  
Gene Promoter ◽  
Synergetic Effects ◽  
Glaa Gene

Regulation of the glaA gene of Aspergillus niger

1990 ◽  
Vol 18 (6) ◽  
pp. 537-545 ◽  
Author(s):  
Timothy Fowler ◽  
Randy M. Berka ◽  
Michael Ward
Keyword(s):  
Aspergillus Niger ◽  
Glaa Gene

scholarly journals Engineering cofactor metabolism for improved protein and glucoamylase production in Aspergillus niger

2020 ◽  
Author(s):  
Yu-fei Sui ◽  
Tabea Schütze ◽  
Li-Ming Ouyang ◽  
Hong-zhong Lu ◽  
Peng Liu ◽  
...  
Keyword(s):  
Metabolic Engineering ◽  
Aspergillus Niger ◽  
Gene Copy ◽  
Cell Factory ◽  
Flux Analysis ◽  
Cofactor Engineering ◽  
Nadph Regeneration ◽  
Gene Copies ◽  
Glaa Gene ◽  
Glucoamylase Production

Abstract Background: Nicotinamide adenine dinucleotide phosphate (NADPH) is an important cofactor ensuring intracellular redox balance, anabolism and cell growth in all living systems. Our recent multi-omics analyses of glucoamylase (GlaA) biosynthesis in the filamentous fungal cell factory Aspergillus niger indicated that low availability of NADPH might be a limiting factor for GlaA overproduction. Results: We thus employed the Design-Build-Test-Learn cycle for metabolic engineering to identify and prioritize effective cofactor engineering strategies for GlaA overproduction. Based on available metabolomics and 13C metabolic flux analysis data, we individually overexpressed seven predicted genes encoding NADPH regeneration enzymes under the control of Tet-on gene switch in two A. niger recipient strains, one carrying a single and one carrying seven glaA gene copies, respectively, to test their individual effects on GlaA overproduction. Both strains were selected to understand if a strong pull towards glaA biosynthesis (seven gene copies) mandates a higher NADPH supply compared to the native condition (one gene copy). Detailed analysis of all 14 strains cultivated in shake flask cultures uncovered that overexpression of the gsdA gene (glucose 6-phosphate dehydrogenase), gndA gene (6-phosphogluconate dehydrogenase) and maeA gene (NADP-dependent malic enzyme) supported GlaA production on a subtle (10%) but significant level in the background strain carrying seven glaA gene copies. We thus performed maltose-limited chemostat cultures combining metabolome analysis for these three isolates to characterize metabolic-level fluctuations caused by cofactor engineering. In these cultures, overexpression of either the gndA or maeA gene increased the intracellular NADPH pool by 45% and 66%, and the yield of GlaA by 65% and 30%, respectively. In contrast, overexpression of the gsdA gene had a negative effect on both total protein and glucoamylase production. Conclusions: This data suggests for the first time that increased NADPH availability can indeed underpin protein and especially GlaA production in strains where a strong pull towards GlaA biosynthesis exists. This data also indicates that the highest impact on GlaA production can be engineered on a genetic level by increasing the flux through the reverse TCA cycle ( maeA gene) followed by engineering the flux through the pentose phosphate pathway ( gndA gene). We thus propose that NADPH cofactor engineering is indeed a valid strategy for metabolic engineering of A. niger to improve GlaA production, a strategy which is certainly also applicable to the rational design of other microbial cell factories.

scholarly journals Growth and product formation in chemostat and recycling cultures by Aspergillus niger N402 and a glucoamylase overproducing transformant, provided with multiple copies of the glaA gene

1993 ◽  
Vol 139 (11) ◽  
pp. 2801-2810 ◽  
Author(s):  
J. M. Schrickx ◽  
A. S. Krave ◽  
J. C. Verdoes ◽  
C. A. M. J. J. van den Hondel ◽  
A. H. Stouthamer ◽  
...  
Keyword(s):  
Aspergillus Niger ◽  
Product Formation ◽  
Multiple Copies ◽  
Glaa Gene

scholarly journals Molecular Characterization and Expression of a Phytase Gene from the Thermophilic Fungus Thermomyces lanuginosus

1998 ◽  
Vol 64 (11) ◽  
pp. 4423-4427 ◽  
Author(s):  
Randy M. Berka ◽  
Michael W. Rey ◽  
Kimberly M. Brown ◽  
Tony Byun ◽  
Alan V. Klotz
Keyword(s):  
Aspergillus Niger ◽  
Transcriptional Control ◽  
Specific Activity ◽  
Gene Promoter ◽  
Catalytic Efficiency ◽  
Thermophilic Fungus ◽  
Thermomyces Lanuginosus ◽  
Gene Encoding ◽  
Putative Signal Peptide ◽  
Ph Range

ABSTRACT The phyA gene encoding an extracellular phytase from the thermophilic fungus Thermomyces lanuginosus was cloned and heterologously expressed, and the recombinant gene product was biochemically characterized. The phyA gene encodes a primary translation product (PhyA) of 475 amino acids (aa) which includes a putative signal peptide (23 aa) and propeptide (10 aa). The deduced amino acid sequence of PhyA has limited sequence identity (ca. 47%) with Aspergillus niger phytase. The phyAgene was inserted into an expression vector under transcriptional control of the Fusarium oxysporum trypsin gene promoter and used to transform a Fusarium venenatum recipient strain. The secreted recombinant phytase protein was enzymatically active between pHs 3 and 7.5, with a specific activity of 110 μmol of inorganic phosphate released per min per mg of protein at pH 6 and 37°C. The Thermomyces phytase retained activity at assay temperatures up to 75°C and demonstrated superior catalytic efficiency to any known fungal phytase at 65°C (the temperature optimum). Comparison of this new Thermomyces catalyst with the well-known Aspergillus niger phytase reveals other favorable properties for the enzyme derived from the thermophilic gene donor, including catalytic activity over an expanded pH range.

614: Polymorphisms in the E-Cadherin (CDH1) Gene Promoter and the Risk of Bladder Cancer

2006 ◽  
Vol 175 (4S) ◽  
pp. 198-198
Author(s):  
Lambertus A. Kiemeney ◽  
Kjeld P. Van Houwelingen ◽  
Manon Bogaerts ◽  
J. Alfred Witjes ◽  
Dorine W. Swinkels ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Gene Promoter ◽  
Cdh1 Gene ◽  
E Cadherin

132: Association of MMP-1 Gene Promoter Polymorphism with the Risk of Prostate Cancer and MMP-1 Expression in Radical Prostatectomy Specimens

2006 ◽  
Vol 175 (4S) ◽  
pp. 42-42
Author(s):  
Norihiko Tsuchiya ◽  
Shintaro Narita ◽  
Teruaki Kumazawa ◽  
Yohei Horikawa ◽  
Hideaki Kakinuma ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Radical Prostatectomy ◽  
Gene Promoter ◽  
Promoter Polymorphism ◽  
Gene Promoter Polymorphism
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