Synchrotron radiation‐based microcomputed tomography for three‐dimensional growth analysis of <i>Aspergillus niger</i> pellets

Müller, Henri; Deffur, Charlotte; Schmideder, Stefan; Barthel, Lars; Friedrich, Tiaan; Mirlach, Lukas; Hammel, Jörg U.; Meyer, Vera; Briesen, Heiko

doi:10.1002/bit.28506

Cited by 8 publications

(2 citation statements)

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“…The image processing pipeline was established in Matlab (Version 2022a) and contained the following parts: (1) the segmentation of hemp shive substrate and (2) the segmentation of fungal hyphae. The preprocessing of the CT-data including the reconstruction of the 3D images and removal of the sample holder based on the methods described in Müller et al 2023 [ 28 ]). Before the segmentation of hemp shives (1), a non-local-mean filter was used (MATLAB function imfiltnlm) to reduce noise in the background for each slice of the 3D image data.…”

Section: Methodsmentioning

confidence: 99%

Mechanical, physical and thermal properties of composite materials produced with the basidiomycete Fomes fomentarius

Schmidt,

Freidank-Pohl,

Zillessen

et al. 2023

Fungal Biol Biotechnol

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Background To achieve climate neutrality, fundamentally new concepts of circularity need to be implemented by the building sector as it contributes to 40% of anthropogenic CO2 emission. Fungal biotechnology can make a significant contribution here and help eliminate fossil dependency for building material production. Recently, we have shown that the medicinal polypore Fomes fomentarius feeds well on renewable lignocellulosic biomass and produces composite materials that could potentially replace fossil fuel-based expanded polystyrene as insulation material. Results In this study, we explored the mechanical, physical, and thermal properties of F. fomentarius-based composite materials in more detail and determined key performance parameters that are important to evaluate the usability of F. fomentarius-based composite materials in the construction sector. These parameters were determined according to European standards and included compressive strength, modulus of elasticity, thermal conductivity, water vapour permeability, and flammability of uncompressed composites as well as flexural strength, transverse tensile strength, and water absorption capacity of heat-pressed composites, among others. We could show that uncompressed composites obtained from F. fomentarius and hemp shives display a thermal conductivity of 0.044 W (m K)−1 which is in the range of natural organic fibres. A water vapour permeability of 1.72 and classification into flammability class B1 clearly surpasses fossil-based insulation materials including expanded polystyrene and polyurethane. We could furthermore show that heat-pressing can be used to reliably generate stiff and firm particleboards that have the potential to replace current wood-based particleboards that contain synthetic additives. X-ray microcomputed tomography finally visualized for the first time the growth of hyphae of F. fomentarius on and into the hemp shive substrates and generated high-resolution images of the microstructure of F. fomentarius-based composites. Conclusion This study demonstrates that fungal-based composites produced with F. fomentarius partially meet or even exceed key performance parameters of currently used fossil fuel-based insulation materials and can also be used to replace particleboards.

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Section: Methodsmentioning

confidence: 99%

Mechanical, physical and thermal properties of composite materials produced with the basidiomycete Fomes fomentarius

Schmidt,

Freidank-Pohl,

Zillessen

et al. 2023

Fungal Biol Biotechnol

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“…3D imaging was conducted at the Deutsches Elektronen Synchrotron (DESY) using the microtomography beamline P05 of PETRA III in Hamburg, Germany as previously described [ 29 – 31 ]. For quantification, images were analysed as previously described [ 47 ].…”

Section: Phylogenetic Analysismentioning

confidence: 99%

Breaking down barriers: comprehensive functional analysis of the Aspergillus niger chitin synthase repertoire

Barthel,

Cairns,

Duda

et al. 2024

Fungal Biol Biotechnol

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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.

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3D imaging and analysis to unveil the impact of microparticles on the pellet morphology of filamentous fungi

Dinius,

Müller,

Kellhammer

et al. 2024

Biotech & Bioengineering

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Controlling the morphology of filamentous fungi is crucial to improve the performance of fungal bioprocesses. Microparticle‐enhanced cultivation (MPEC) increases productivity, most likely by changing the fungal morphology. However, due to a lack of appropriate methods, the exact impact of the added microparticles on the structural development of fungal pellets is mostly unexplored. In this study synchrotron radiation‐based microcomputed tomography and three‐dimensional (3D) image analysis were applied to unveil the detailed 3D incorporation of glass microparticles in nondestructed pellets of Aspergillus niger from MPEC. The developed method enabled the 3D analysis based on 375 pellets from various MPEC experiments. The total and locally resolved volume fractions of glass microparticles and hyphae were quantified for the first time. At increasing microparticle concentrations in the culture medium, pellets with lower hyphal fraction were obtained. However, the total volume of incorporated glass microparticles within the pellets did not necessarily increase. Furthermore, larger microparticles were less effective than smaller ones in reducing pellet density. However, the total volume of incorporated glass was larger for large microparticles. In addition, analysis of MPEC pellets from different times of cultivation indicated that spore agglomeration is decisive for the development of MPEC pellets. The developed 3D morphometric analysis method and the presented results will promote the general understanding and further development of MPEC for industrial application.

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Synchrotron radiation‐based microcomputed tomography for three‐dimensional growth analysis of Aspergillus niger pellets

Cited by 8 publications

References 69 publications

Mechanical, physical and thermal properties of composite materials produced with the basidiomycete Fomes fomentarius

Mechanical, physical and thermal properties of composite materials produced with the basidiomycete Fomes fomentarius

Breaking down barriers: comprehensive functional analysis of the Aspergillus niger chitin synthase repertoire

3D imaging and analysis to unveil the impact of microparticles on the pellet morphology of filamentous fungi

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