ArticleApplied and environmental microbiology2002
Silencing of the aspergillopepsin B (pepB) gene of Aspergillus awamori by antisense RNA expression or protease removal by gene disruption results in a large increase in thaumatin production.
Article in Applied and environmental microbiology, 2002. 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, 50 citations in OpenAlex.
- Exploration of the Strategy for Improving the Expression of Heterologous Sweet Protein Monellin inJournal of fungi (Basel, Switzerland) · 2023Article
- Study of protease activity from Aspergillus awamori INCQS2B.361U2/1 extracellular fraction and modification of culture medium composition to isolate a novel aspartic protease.Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] · 2022Article
- Proteolytic analysis of Trichoderma reesei in celluase-inducing condition reveals a role for trichodermapepsin (TrAsP) in cellulase production.Journal of industrial microbiology & biotechnology · 2019Article
- Forced Recycling of an AMA1-Based Genome-Editing Plasmid Allows for Efficient Multiple Gene Deletion/Integration in the Industrial Filamentous FungusApplied and environmental microbiology · 2019Article
- Engineered Microorganisms for the Production of Food Additives Approved by the European Union-A Systematic Analysis.Frontiers in microbiology · 2018Review
- Enabling low cost biopharmaceuticals: high level interferon alpha-2b production in Trichoderma reesei.Microbial cell factories · 2016Article
- Aspergillus as a multi-purpose cell factory: current status and perspectives.Biotechnology letters · 2011Review
- A comparison of methods for successful triggering of gene silencing in Coprinus cinereus.Molecular biotechnology · 2007Article
- Protein kinase C (PkcA) of Aspergillus nidulans is involved in penicillin production.Applied and environmental microbiology · 2006Article
- Disruption of the subtilase gene, albin1, in Ophiostoma piliferum.Applied and environmental microbiology · 2004Article
- Antisense RNA: the new favorite in genetic research.Journal of Zhejiang University. Science. BReview
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Aspergillopepsin B was identified in culture broths of Aspergillus awamori by in situ detection of its proteolytic activity and by immunodetection with anti-aspergillopepsin B antibodies. Severe thaumatin degradation was observed after in vitro treatment of thaumatin with purified aspergillopepsin B. The pepB gene encoding aspergillopepsin B of A. awamori was cloned and characterized. It is located in chromosome IV of A. awamori, as shown by pulsed-field gel electrophoresis, and encodes a protein of 282 amino acids with high similarity to the aspergillopepsin B of Aspergillus niger var. macrosporus. The pepB gene is expressed at high rates as a monocistronic 1.0-kb transcript in media with casein at acidic pH values. An antisense cassette constructed by inserting the pepB gene in the antisense orientation downstream from the gpdA promoter resulted in a good level of antisense mRNA, as shown by reverse transcription-PCR. Partial silencing of the pepB gene by the antisense mRNA resulted in a 31% increase in thaumatin yield. However, significant residual degradation of thaumatin still occurred. To completely remove aspergillopepsin B, the pepB gene was deleted by double crossover. Two of the selected transformants lacked the endogenous pepB gene and did not form aspergillopepsin B. Thaumatin yields increased by between 45% in transformant APB 7/25 and 125% in transformant 7/36 with respect to the parental strain. Reduction of proteolytic degradation by gene silencing with antisense mRNA or total removal of the aspergillopepsin B by directed gene deletion was a very useful method for improving thaumatin production in A. awamori.
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