Cell Wall Integrity and Its Industrial Applications in Filamentous Fungi

Yoshimi, Akira; Miyazawa, Ken; Kawauchi, Moriyuki; Abe, Keietsu

doi:10.3390/jof8050435

Cited by 16 publications

(6 citation statements)

References 158 publications

(249 reference statements)

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“…Fungi exhibit a cell wall integrity response; this is a signaling pathway activated by cell wall stress or damage involving fungal-specific kinases 53 and is the basic survival mechanism. The cell wall integrity pathway is involved in the response to osmotic stress 54 , fungal morphogenesis, and fungal pathogenicity.…”

Section: Discussionmentioning

confidence: 99%

In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi

Pajić,

Stevanović,

Todorović

et al. 2024

Microsyst Nanoeng

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Studying the membrane physiology of filamentous fungi is key to understanding their interactions with the environment and crucial for developing new therapeutic strategies for disease-causing pathogens. However, their plasma membrane has been inaccessible for a micron-sized patch-clamp pipette for pA current recordings due to the rigid chitinous cell wall. Here, we report the first femtosecond IR laser nanosurgery of the cell wall of the filamentous fungi, which enabled patch-clamp measurements on protoplasts released from hyphae. A reproducible and highly precise (diffraction-limited, submicron resolution) method for obtaining viable released protoplasts was developed. Protoplast release from the nanosurgery-generated incisions in the cell wall was achieved from different regions of the hyphae. The plasma membrane of the obtained protoplasts formed tight and high-resistance (GΩ) contacts with the recording pipette. The entire nanosurgical procedure followed by the patch-clamp technique could be completed in less than 1 hour. Compared to previous studies using heterologously expressed channels, this technique provides the opportunity to identify new ionic currents and to study the properties of the ion channels in the protoplasts of filamentous fungi in their native environment.

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Section: Discussionmentioning

confidence: 99%

In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi

Pajić,

Stevanović,

Todorović

et al. 2024

Microsyst Nanoeng

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show abstract

“…Fungi have cell wall integrity response, a signaling pathway activated by cell wall stress or damage and involving fungal-speci c kinases 52 , which is the basic survival mechanism. The cell wall integrity pathway is involved in the response to osmotic stress 53 , fungal morphogenesis, and fungal pathogenicity.…”

Section: Discussionmentioning

confidence: 99%

In vivo femtosecond laser nanosurgery of cell wall enables patch-clamp on filamentous fungi

Pajić,

Stevanović,

Todorović

et al. 2023

Preprint

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Studying the membrane physiology of filamentous fungi is key to understanding their interactions with the environment and crucial for developing new therapeutic strategies for disease-causing pathogens. However, their plasma membrane has been inaccessible for micron sized patch-clamp pipette for pA currents recordings due to the rigid chitinous cell wall. We report here the first femtosecond IR laser nanosurgery of the cell wall of filamentous fungi, enabling patch-clamp measurements on protoplasts released from hyphae. A reproducible and highly precise (diffraction limited, sub-micron resolution) method for obtaining viable released protoplasts was developed. Protoplast release from the nanosurgery-generated incisions in cell wall was achieved from different regions of the hyphae. Plasma membrane of obtained protoplasts formed tight and high-resistance (GΩ) contacts with the recording pipette. The entire nanosurgical procedure followed by patch-clamp technique can be completed in less than 1 hour. Compared to previous studies using heterologously expressed channels, this technique provides the opportunity to identify new ionic currents and to study the properties of ion channels in protoplasts of filamentous fungi in their native environment.

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“…These characteristics were the presence of hydroxyls at C3 and C7 positions of flavonoids, and the O ‐ and C ‐glycosylation, methylation, and acetylation of OH groups (Jakimiuk et al, 2022 ). This target‐based drug discovery strategy was also used to identify compounds that inhibit proteins of the ergosterol biosynthesis pathway, proteins involved in cell‐wall construction, or those involved in signalling systems (Ahmad et al, 2010 ; Yoshimi et al, 2022 ).…”

Section: The Target‐based Drug‐discovery Strategymentioning

confidence: 99%

Modes‐of‐action of antifungal compounds: Stressors and (target‐site‐specific) toxins, toxicants, or toxin–stressors

Noël

Hallsworth

Gelhaye

et al. 2023

Microbial Biotechnology

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Fungi and antifungal compounds are relevant to the United Nation's Sustainable Development Goals. However, the modes‐of‐action of antifungals—whether they are naturally occurring substances or anthropogenic fungicides—are often unknown or are misallocated in terms of their mechanistic category. Here, we consider the most effective approaches to identifying whether antifungal substances are cellular stressors, toxins/toxicants (that are target‐site‐specific), or have a hybrid mode‐of‐action as toxin–stressors (that induce cellular stress yet are target‐site‐specific). This newly described ‘toxin–stressor’ category includes some photosensitisers that target the cell membrane and, once activated by light or ultraviolet radiation, cause oxidative damage. We provide a glossary of terms and a diagrammatic representation of diverse types of stressors, toxic substances, and toxin–stressors, a classification that is pertinent to inhibitory substances not only for fungi but for all types of cellular life. A decision‐tree approach can also be used to help differentiate toxic substances from cellular stressors (Curr Opin Biotechnol 2015 33: 228–259). For compounds that target specific sites in the cell, we evaluate the relative merits of using metabolite analyses, chemical genetics, chemoproteomics, transcriptomics, and the target‐based drug‐discovery approach (based on that used in pharmaceutical research), focusing on both ascomycete models and the less‐studied basidiomycete fungi. Chemical genetic methods to elucidate modes‐of‐action currently have limited application for fungi where molecular tools are not yet available; we discuss ways to circumvent this bottleneck. We also discuss ecologically commonplace scenarios in which multiple substances act to limit the functionality of the fungal cell and a number of as‐yet‐unresolved questions about the modes‐of‐action of antifungal compounds pertaining to the Sustainable Development Goals.

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Cell Wall Integrity and Its Industrial Applications in Filamentous Fungi

Cited by 16 publications

References 158 publications

In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi

In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi

In vivo femtosecond laser nanosurgery of cell wall enables patch-clamp on filamentous fungi

Modes‐of‐action of antifungal compounds: Stressors and (target‐site‐specific) toxins, toxicants, or toxin–stressors

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