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Mycoremediation of copper-azole wood preservatives by Phanerochaete chrysosporium with a focus on resistance mechanisms and azole detoxification

ABSTRACT: Mycoremediation of copper-azole wood preservatives by Phanerochaete chrysosporium with a focus on resistance mechanisms and azole detoxification

PROVIDER: PRJNA1197371 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR31694867_1.fastq.gz	Fastqsanger.gz
	SRR31694867_2.fastq.gz	Fastqsanger.gz
	SRR31694868_1.fastq.gz	Fastqsanger.gz
	SRR31694868_2.fastq.gz	Fastqsanger.gz
	SRR31694869_1.fastq.gz	Fastqsanger.gz

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Mycoremediation of copper-azole wood preservatives by Phanerochaete chrysosporium with a focus on resistance mechanisms and azole detoxification

Project description:Copper-azole based formulations were widely used against fungal decay for preservation of wood timbers. While efficient for wood protection, leaching of both copper and azole within the environment has deleterious impact on soils and surface waters. Moreover, no recycling process is currently available for these wood waste. By taking advantage of the fact that certain fungi have the capacity to cope with these active compounds, we propose that some ligninolytic fungi could be good biocatalysts for detoxifying copper-azole formulations. Using the white-rot fungus Phanerochaete chrysosporium as a model, we demonstrated that the main strategies implemented by this fungus to counteract the antifungal effect of the product, in particular azoles, are the modulation of the lipids and sterols content, the maintenance of DNA integrity, detoxification of azoles by extracellular degradation likely through the Fenton chemistry, biosorption at the cell wall, efflux, and intracellular detoxification by the three-step detoxification pathway. By using comparative transcriptomics between a copper-azole formulation and a copper-quaternary ammonium formulation, we manage to identify genes specifically involved in azole resistance and detoxification within this complex system, opening perspectives for both the identification of key molecular players that could be the targets for developing long-term acquired antifungal resistance mechanisms, and the management of azoles residues by mycoremediation processes.

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Project description:The transcriptome of Phanerochaete chrysosporium control mycelium was compared to the transcriptome of mycelium grown on oak acetonic extractives containing medium. The array probes were designed from gene models taken from the Joint Genome Institute (JGI, Department of Energy) Phanerochaete chrysosporium genome sequence version 1. The aim of this study was to determine gene expression changes in Phanerochaete chrysosporium grown on oak extract with a special focus on detoxification systems.

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Project description:Using whole genome microarrays based on the annotated genomes of Phanerochaete chrysosporium, we monitored the changes in its transcriptomes relevant to cell wall degradation during growth on three chemically distinct Populus trichocarpa (poplar) wood substrates. Results of this study are sumbitted for review in Biotechnology for Biofuels

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Aprepitant, an antiemetic agent, displays potent synergistic interactions with azole drugs against emerging multidrug-resistant Candida auris

Project description:Combination therapies can be a promising tool to augment the antifungal activity of azole drugs against resistant Candida species. Here, we report the interaction between aprepitant, an antiemetic agent, and azole drugs against different Candida species including the emerging multidrug-resistant C. auris. Particularly, aprepitant enhanced the antifungal activity of itraconazole against C. auris by reducing its minimum inhibitory concentration (MIC) by 2-8 folds. Using Caenorhabditis elegans as an in vivo infection model, the aprepitant/itraconazole combination significantly prolonged the survival of the infected nematodes by ~90% and reduced the fungal burden by ~92% relative to the untreated control. Interestingly, the aprepitant/itraconazole combination exerted a potent fungicidal activity against both planktonic and adherent C. auris biofilms. Further, aprepitant/itraconazole displayed broad-spectrum synergistic interactions against other medically important Candida species including C. albicans, C. krusie, C. tropicalis, and C. parapsilosis (ƩFICI ranged from 0.08 to 031). Comparative transcriptomic profiling indicated aprepitant/itraconazole interferes significantly with metal ions homeostasis and compromises the ROS (reactive oxygen species) detoxification ability of C. auris. This study presents aprepitant as a novel, potent and broad-spectrum azole chemosensitizing agent that warrants further investigation.

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UPC2 Is universally essential for Azole antifungal resistance in Candida albicans

Project description:In Candida albicans, the transcription factor Upc2 is central to the regulation of ergosterol biosynthesis. UPC2-activating mutations contribute to azole resistance, whereas disruption increases azole susceptibility. In the present study, we investigated the relationship of UPC2 to fluconazole susceptibility, particularly in azole-resistant strains. In addition to the reduced fluconazole MIC previously observed with UPC2 disruption, we observed a lower minimum fungicidal concentration (MFC) for a upc2Δ/Δ mutant than for its azole-susceptible parent, SC5314. Moreover, the upc2Δ/Δ mutant was unable to grow on a solid medium containing 10 µg/ml fluconazole and exhibited increased susceptibility and a clear zone of inhibition by Etest. Time-kill analysis showed higher fungistatic activity against the upc2Δ/Δ mutant than against SC5314. UPC2 disruption in strains carrying specific resistance mutations also resulted in reduced MICs and MFCs. UPC2 disruption in a highly azole resistant clinical isolate containing multiple resistance mechanisms likewise resulted in a reduced MIC and MFC. This mutant was unable to grow on a solid medium containing 10 µg/ml fluconazole and exhibited increased susceptibility and a clear zone of inhibition by Etest. Time-kill analysis showed increased fungistatic activity against the upc2Δ/Δ mutant in the resistant background. Microarray analysis showed attenuated induction by fluconazole of genes involved in sterol biosynthesis, iron transport, or iron homeostasis in the absence of UPC2. Taken together, these data demonstrate that the UPC2 transcriptional network is universally essential for azole resistance in C. albicans and represents an attractive target for enhancing azole antifungal activity.

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