ArticleBioengineered
Random and direct mutagenesis to enhance protein secretion in Ashbya gossypii.
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Who cites it
13 citing papers in PubMed, 47 citations in OpenAlex.
- Hyperproduction of nattokinase from Bacillus subtilis VIT MS2 using random mutagenesis and statistical optimization through central composite design.BMC microbiology · 2025Article
- Creating a novel genetic diversity ofHeliyon · 2024Article
- Unveiling a classical mutant in the context of the GH3 β-glucosidase family in Neurospora crassa.AMB Express · 2024Article
- Strategies for the Development of Industrial Fungal Producing Strains.Journal of fungi (Basel, Switzerland) · 2023Review
- Heterologous protein production in filamentous fungi.Applied microbiology and biotechnology · 2023Review
- Plasma Promotes Fungal Cellulase Production by Regulating the Levels of Intracellular NO and CaInternational journal of molecular sciences · 2022Article
- Keratinolytic activity of3 Biotech · 2019Article
- New biotechnological applications for Ashbya gossypii: Challenges and perspectives.Bioengineered · 2017Review
- Investigation of protein secretion and secretion stress in Ashbya gossypii.BMC genomics · 2014Article
- Improving industrial yeast strains: exploiting natural and artificial diversity.FEMS microbiology reviews · 2014Review
- Molecular and functional characterization of an invertase secreted by Ashbya gossypii.Molecular biotechnology · 2014Article
- Strain design of Ashbya gossypii for single-cell oil production.Applied and environmental microbiology · 2014Article
- Making recombinant proteins in filamentous fungi- are we expecting too much?Frontiers in microbiology · 2014Review
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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
To improve the general secretion ability of the biotechnologically relevant fungus Ashbya gossypii, random mutagenesis with ethyl methane sulfonate (EMS) was performed. The selection and screening strategy followed revealed mutants with improved secretion of heterologous Trichoderma reesei endoglucanase I (EGI), native α-amylase and/or native β-glucosidase. One mutant, S436, presented 1.4- to 2-fold increases in all extracellular enzymatic activities measured, when compared with the parent strain, pointing to a global improvement in protein secretion. Three other mutants exhibited 2- to 3-fold improvements in only one (S397, B390) or two (S466) of the measured activities. A targeted genetic approach was also followed. Two homologs of the Saccharomyces cerevisiae GAS1, AgGAS1A (AGL351W) and AgGAS1B (AGL352W), were deleted from the A. gossypii genome. For both copies deletion, a new antibiotic marker cassette conferring resistance to phleomycin, BLE3, was constructed. GAS1 encodes an β-1,3-glucanosyltransglycosylase involved in cell wall assembly. Higher permeability of the cell wall was expected to increase the protein secretion capacity. However, total protein secreted to culture supernatants and secreted EGI activity did not increase in the Aggas1AΔ mutants. Deletion of the AgGAS1B copy affected cellular morphology and resulted in severe retardation of growth, similarly to what has been reported for GAS1-defficient yeast. Thus, secretion could not be tested in these mutants.
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