Breaking down barriers: comprehensive functional analysis of the Aspergillus niger chitin synthase repertoire
Abstract Background Members of the fungal kingdom are heterotrophic eukaryotes encased in a chitin containing cell wall. This polymer is vital for cell wall stiffness and, ultimately, cell shape. Most fungal genomes contain numerous putative chitin synthase encoding genes. However, systematic functi...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2024-03-01
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| Series: | Fungal Biology and Biotechnology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s40694-024-00172-7 |
| _version_ | 1797259599151104000 |
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| author | Lars Barthel Timothy Cairns Sven Duda Henri Müller Birgit Dobbert Sascha Jung Heiko Briesen Vera Meyer |
| author_facet | Lars Barthel Timothy Cairns Sven Duda Henri Müller Birgit Dobbert Sascha Jung Heiko Briesen Vera Meyer |
| author_sort | Lars Barthel |
| collection | DOAJ |
| description | Abstract Background Members of the fungal kingdom are heterotrophic eukaryotes encased in a chitin containing cell wall. This polymer is vital for cell wall stiffness and, ultimately, cell shape. Most fungal genomes contain numerous putative chitin synthase encoding genes. However, systematic functional analysis of the full chitin synthase catalogue in a given species is rare. This greatly limits fundamental understanding and potential applications of manipulating chitin synthesis across the fungal kingdom. Results In this study, we conducted in silico profiling and subsequently deleted all predicted chitin synthase encoding genes in the multipurpose cell factory Aspergillus niger. Phylogenetic analysis suggested nine chitin synthases evolved as three distinct groups. Transcript profiling and co-expression network construction revealed remarkably independent expression, strongly supporting specific role(s) for the respective chitin synthases. Deletion mutants confirmed all genes were dispensable for germination, yet impacted colony spore titres, chitin content at hyphal septa, and internal architecture of submerged fungal pellets. We were also able to assign specific roles to individual chitin synthases, including those impacting colony radial growth rates (ChsE, ChsF), lateral cell wall chitin content (CsmA), chemical genetic interactions with a secreted antifungal protein (CsmA, CsmB, ChsE, ChsF), resistance to therapeutics (ChsE), and those that modulated pellet diameter in liquid culture (ChsA, ChsB). From an applied perspective, we show chsF deletion increases total protein in culture supernatant over threefold compared to the control strain, indicating engineering filamentous fungal chitin content is a high priority yet underexplored strategy for strain optimization. Conclusion This study has conducted extensive analysis for the full chitin synthase encoding gene repertoire of A. niger. For the first time we reveal both redundant and non-redundant functional roles of chitin synthases in this fungus. Our data shed light on the complex, multifaceted, and dynamic role of chitin in fungal growth, morphology, survival, and secretion, thus improving fundamental understanding and opening new avenues for biotechnological applications in fungi. |
| first_indexed | 2024-04-24T23:11:59Z |
| format | Article |
| id | doaj.art-cd8a2b8eb5284505b28bcf94d62217cb |
| institution | Directory Open Access Journal |
| issn | 2054-3085 |
| language | English |
| last_indexed | 2024-04-24T23:11:59Z |
| publishDate | 2024-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Fungal Biology and Biotechnology |
| spelling | doaj.art-cd8a2b8eb5284505b28bcf94d62217cb2024-03-17T12:12:45ZengBMCFungal Biology and Biotechnology2054-30852024-03-0111111710.1186/s40694-024-00172-7Breaking down barriers: comprehensive functional analysis of the Aspergillus niger chitin synthase repertoireLars Barthel0Timothy Cairns1Sven Duda2Henri Müller3Birgit Dobbert4Sascha Jung5Heiko Briesen6Vera Meyer7Chair of Applied and Molecular Microbiology, Institute of Biotechnology, Technische Universität BerlinChair of Applied and Molecular Microbiology, Institute of Biotechnology, Technische Universität BerlinChair of Applied and Molecular Microbiology, Institute of Biotechnology, Technische Universität BerlinSchool of Life Sciences Weihenstephan, Chair of Process Systems Engineering, Technical University of MunichChair of Applied and Molecular Microbiology, Institute of Biotechnology, Technische Universität BerlinChair of Applied and Molecular Microbiology, Institute of Biotechnology, Technische Universität BerlinSchool of Life Sciences Weihenstephan, Chair of Process Systems Engineering, Technical University of MunichChair of Applied and Molecular Microbiology, Institute of Biotechnology, Technische Universität BerlinAbstract Background Members of the fungal kingdom are heterotrophic eukaryotes encased in a chitin containing cell wall. This polymer is vital for cell wall stiffness and, ultimately, cell shape. Most fungal genomes contain numerous putative chitin synthase encoding genes. However, systematic functional analysis of the full chitin synthase catalogue in a given species is rare. This greatly limits fundamental understanding and potential applications of manipulating chitin synthesis across the fungal kingdom. Results In this study, we conducted in silico profiling and subsequently deleted all predicted chitin synthase encoding genes in the multipurpose cell factory Aspergillus niger. Phylogenetic analysis suggested nine chitin synthases evolved as three distinct groups. Transcript profiling and co-expression network construction revealed remarkably independent expression, strongly supporting specific role(s) for the respective chitin synthases. Deletion mutants confirmed all genes were dispensable for germination, yet impacted colony spore titres, chitin content at hyphal septa, and internal architecture of submerged fungal pellets. We were also able to assign specific roles to individual chitin synthases, including those impacting colony radial growth rates (ChsE, ChsF), lateral cell wall chitin content (CsmA), chemical genetic interactions with a secreted antifungal protein (CsmA, CsmB, ChsE, ChsF), resistance to therapeutics (ChsE), and those that modulated pellet diameter in liquid culture (ChsA, ChsB). From an applied perspective, we show chsF deletion increases total protein in culture supernatant over threefold compared to the control strain, indicating engineering filamentous fungal chitin content is a high priority yet underexplored strategy for strain optimization. Conclusion This study has conducted extensive analysis for the full chitin synthase encoding gene repertoire of A. niger. For the first time we reveal both redundant and non-redundant functional roles of chitin synthases in this fungus. Our data shed light on the complex, multifaceted, and dynamic role of chitin in fungal growth, morphology, survival, and secretion, thus improving fundamental understanding and opening new avenues for biotechnological applications in fungi.https://doi.org/10.1186/s40694-024-00172-7Aspergillus nigerChitinCell wallµCTProtein secretionMacromorphology |
| spellingShingle | Lars Barthel Timothy Cairns Sven Duda Henri Müller Birgit Dobbert Sascha Jung Heiko Briesen Vera Meyer Breaking down barriers: comprehensive functional analysis of the Aspergillus niger chitin synthase repertoire Fungal Biology and Biotechnology Aspergillus niger Chitin Cell wall µCT Protein secretion Macromorphology |
| title | Breaking down barriers: comprehensive functional analysis of the Aspergillus niger chitin synthase repertoire |
| title_full | Breaking down barriers: comprehensive functional analysis of the Aspergillus niger chitin synthase repertoire |
| title_fullStr | Breaking down barriers: comprehensive functional analysis of the Aspergillus niger chitin synthase repertoire |
| title_full_unstemmed | Breaking down barriers: comprehensive functional analysis of the Aspergillus niger chitin synthase repertoire |
| title_short | Breaking down barriers: comprehensive functional analysis of the Aspergillus niger chitin synthase repertoire |
| title_sort | breaking down barriers comprehensive functional analysis of the aspergillus niger chitin synthase repertoire |
| topic | Aspergillus niger Chitin Cell wall µCT Protein secretion Macromorphology |
| url | https://doi.org/10.1186/s40694-024-00172-7 |
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