Enzymatic Adaptation of <i>Podospora anserina</i> to Different Plant Biomass Provides Leads to Optimized Commercial Enzyme Cocktails

Benocci, Tiziano; Daly, Paul; Pontes, María Victoria Aguilar; Lail, Kathleen; Wang, Mei; Lipzen, Anna; Ng, Vivian; Grigoriev, Igor V.

doi:10.1002/biot.201800185

Cited by 11 publications

(9 citation statements)

References 31 publications

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“…Media and growth conditions for P. anserina were based on previously described procedures [21,60]. Waterinsoluble wheat glucuronoarabinoxylan (GAX, Megazyme, Wicklow, Ireland, 100 mg) and GAX (100 mg) combined with wheat straw lignin isolate (100 mg) were dispersed in 5 mL M2 minimal medium in 40-mL flasks and autoclaved prior to inoculation.…”

Section: Growth Of Podospora Anserina On Wheat Straw Ligninmentioning

confidence: 99%

Evidence for ligninolytic activity of the ascomycete fungus Podospora anserina

Erven

Kleijn

Patyshakuliyeva

et al. 2020

Biotechnol Biofuels

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Background: The ascomycete fungus Podospora anserina has been appreciated for its targeted carbohydrate-active enzymatic arsenal. As a late colonizer of herbivorous dung, the fungus acts specifically on the more recalcitrant fraction of lignocellulose and this lignin-rich biotope might have resulted in the evolution of ligninolytic activities. However, the lignin-degrading abilities of the fungus have not been demonstrated by chemical analyses at the molecular level and are, thus far, solely based on genome and secretome predictions. To evaluate whether P. anserina might provide a novel source of lignin-active enzymes to tap into for potential biotechnological applications, we comprehensively mapped wheat straw lignin during fungal growth and characterized the fungal secretome.Results: Quantitative 13 C lignin internal standard py-GC-MS analysis showed substantial lignin removal during the 7 days of fungal growth (24% w/w), though carbohydrates were preferably targeted (58% w/w removal). Structural characterization of residual lignin by using py-GC-MS and HSQC NMR analyses demonstrated that C α -oxidized substructures significantly increased through fungal action, while intact β-O-4′ aryl ether linkages, p-coumarate and ferulate moieties decreased, albeit to lesser extents than observed for the action of basidiomycetes. Proteomic analysis indicated that the presence of lignin induced considerable changes in the secretome of P. anserina. This was particularly reflected in a strong reduction of cellulases and galactomannanases, while H 2 O 2 -producing enzymes clearly increased. The latter enzymes, together with laccases, were likely involved in the observed ligninolysis. Conclusions:For the first time, we provide unambiguous evidence for the ligninolytic activity of the ascomycete fungus P. anserina and expand the view on its enzymatic repertoire beyond carbohydrate degradation. Our results can be of significance for the development of biological lignin conversion technologies by contributing to the quest for novel lignin-active enzymes and organisms.

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Section: Growth Of Podospora Anserina On Wheat Straw Ligninmentioning

confidence: 99%

Evidence for ligninolytic activity of the ascomycete fungus Podospora anserina

Erven

Kleijn

Patyshakuliyeva

et al. 2020

Biotechnol Biofuels

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“…Even on its preferred natural growth substrate, dung, we detect large variations in the amounts of mycelium and fruiting bodies that the fungus produces. The reasons for this are likely multifactorial and include the secretion of different enzymatic cocktails, as previously demonstrated [29,70,71]. It was shown that the presence of lignin reduces the secretion of cellulases and galactomannanases and increases that of laccases and H 2 O 2 -producing enzymes, which should favor ligninolysis [29].…”

Section: Discussionmentioning

confidence: 85%

Lignin Degradation and Its Use in Signaling Development by the Coprophilous Ascomycete Podospora anserina

Dicko¹,

Ferrari

Tangthirasunun

et al. 2020

JoF

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The filamentous fungus Podospora anserina is a good model to study the breakdown of lignocellulose, owing to its ease of culture and genetical analysis. Here, we show that the fungus is able to use a wide range of lignocellulosic materials as food sources. Using color assays, spectroscopy and pyrolysis–gas chromatography mass spectrometry, we confirm that this ascomycete is able to degrade lignin, primarily by hydrolyzing β–O-4 linkages, which facilitates its nutrient uptake. We show that the limited weight loss that is promoted when attacking Miscanthus giganteus is due to a developmental blockage rather than an inefficiency of its enzymes. Finally, we show that lignin, and, more generally, phenolics, including degradation products of lignin, greatly stimulate the growth and fertility of the fungus in liquid cultures. Analyses of the CATΔΔΔΔΔ mutant lacking all its catalases, pro-oxidants and antioxidants indicate that improved growth and fertility of the fungus is likely caused by augmented reactive oxygen species levels triggered by the presence of phenolics.

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“…Collection of multiple RNA-seq data from various experimental conditions A search for P. anserina in the SRA and BioProject repositories (27,28) returned 44 RNA-seq data from different studies on P. anserina's life cycle (25), adaptation to carbon sources (29), response to bacteria (30) and senescence (31). Because it was generated by the SuperSAGE technology, this last dataset was excluded from the analyses.…”

Section: Resultsmentioning

confidence: 99%

Re-exploiting multiple RNA-seq data to identify transcript variations in Podospora anserina

Lelandais

Remy

Malagnac

et al. 2022

Preprint

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Publicly available RNA-seq datasets are often underused although being helpful to improve functional annotation of eukaryotic genomes. This is especially true for filamentous fungi genomes which structure differs from most well annotated yeast genomes. Podospora anserina is a filamentous fungal model, which genome has been sequenced and annotated in 2008. Still, the current annotation lacks information about cis-regulatory elements, including promoters, transcription starting sites and terminators, which are instrumental to integrate epigenomic features into global gene regulation strategies. Here we took advantage of 37 RNA-seq experiments that were obtained in contrasted developmental and physiological conditions, to complete the functional annotation of P. anserina genome. Out of the 10,800 previously annotated genes, 5'UTR and 3'UTR were defined for 7,554, among which, 3,328 showed differential transcriptional signal starts and/or transcriptional end sites. In addition, alternative splicing events were detected for 2350 genes, mostly due alternative 3'splice site and 1,958 novel transcriptionally active regions (nTARs) in unannotated regions were identified. Our study provides a comprehensive genome-wide functional annotation release of P. anserina genome, including chromatin features, cis-acting elements such as UTRs, alternative splicing events and transcription of non-coding regions. These new findings will likely improve our understanding of gene regulation strategies in compact genomes, such as those of filamentous fungi. Characterization of alternative transcripts and nTARs paves the way to the discovery of putative new genes, alternative peptides or regulatory non-coding RNAs.

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Enzymatic Adaptation of Podospora anserina to Different Plant Biomass Provides Leads to Optimized Commercial Enzyme Cocktails

Cited by 11 publications

References 31 publications

Evidence for ligninolytic activity of the ascomycete fungus Podospora anserina

Evidence for ligninolytic activity of the ascomycete fungus Podospora anserina

Lignin Degradation and Its Use in Signaling Development by the Coprophilous Ascomycete Podospora anserina

Re-exploiting multiple RNA-seq data to identify transcript variations in Podospora anserina

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