ArticleMicrobial cell factories2016
Highly active promoters and native secretion signals for protein production during extremely low growth rates in Aspergillus niger.
Article in Microbial cell factories, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 24 citations in OpenAlex.
- Protein production dynamics and physiological adaptation of recombinant Komagataella phaffii at near-zero growth rates.Microbial cell factories · 2024Article
- The antifungal peptide AnAFP fromFrontiers in microbiology · 2024Article
- Characterising the metabolic rewiring of extremely slow growing Komagataella phaffii.Microbial biotechnology · 2024Article
- The α-(1,3)-glucan synthase gene agsE impacts the secretome of Aspergillus niger.Antonie van Leeuwenhoek · 2023Article
- CRISPR/Cas9-Mediated Multiplexed Genome Editing inJournal of fungi (Basel, Switzerland) · 2023Article
- Protein expression and secretion by filamentous fungi.Journal of biosciences · 2021Review
- Antifungal Peptides of the AFP Family Revisited: Are These Cannibal Toxins?Microorganisms · 2018Review
- Effect of CBM1 and linker region on enzymatic properties of a novel thermostable dimeric GH10 xylanase (Xyn10A) from filamentous fungus Aspergillus fumigatus Z5.AMB Express · 2018Article
- The Promoter Toolbox for Recombinant Gene Expression inFrontiers in bioengineering and biotechnology · 2018Review
- Promoters from the itaconate cluster ofFungal biology and biotechnology · 2017Article
- A Transcriptome Meta-Analysis Proposes Novel Biological Roles for the Antifungal Protein AnAFP in Aspergillus niger.PloS one · 2016Article
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Authors and funding
6 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundThe filamentous ascomycete Aspergillus niger is used in many industrial processes for the production of enzymes and organic acids by batch and fed-batch cultivation. An alternative technique is continuous cultivation, which promises improved yield and optimized pipeline efficiency.
resultsIn this work, we have used perfusion (retentostat) cultivation to validate two promoters that are suitable for A. niger continuous cultivation of industrially relevant products. Firstly, promoters of genes encoding either an antifungal protein (Panafp) or putative hydrophobin (PhfbD) were confirmed as active throughout retentostat culture by assessing mRNA and protein levels using a luciferase (mluc) reporter system. This demonstrated the anafp promoter mediates a high but temporally variable expression profile, whereas the hfbD promoter mediates a semi-constant, moderate-to-high protein expression during retentostat culture. In order to assess whether these promoters were suitable to produce heterologous proteins during retentostat cultivation, the secreted antifungal protein (AFP) from Aspergillus giganteus, which has many potential biotechnological applications, was expressed in A. niger during retentostat cultivation. Additionally, this assay was used to concomitantly validate that native secretion signals encoded in anafp and hfbD genes can be harnessed for secretion of heterologous proteins. Afp mRNA and protein abundance were comparable to luciferase measurements throughout retentostat cultivation, validating the use of Panafp and PhfbD for perfusion cultivation. Finally, a gene encoding the highly commercially relevant thermal hysteresis protein (THP) was expressed in this system, which did not yield detectable protein.
conclusionBoth hfbD and anafp promoters are suitable for production of useful products in A. niger during perfusion cultivation. These findings provide a platform for further optimisations for high production of heterologous proteins with industrial relevance.
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