Open Access
Genomic analysis enlightens agaricales lifestyle evolution and increasing peroxidase diversity
(Oxford University Press, 2021) Ruiz Dueñas, Francisco J.; Barrasa, José M.; Sánchez-García, Marisol; Camarero, Susana; Miyauchi, Shingo; Linde, Dolores; Babiker, Rashid; Drula, Elodie; Ayuso-Fernández, Iván; Pacheco, Remedios; Padilla, Guillermo; Ferreira, Patricia; Barriuso, Jorge; Kellner, Harald; Castanera Andrés, Raúl; Alfaro Sánchez, Manuel; Ramírez Nasto, Lucía; Pisabarro de Lucas, Gerardo; Riley, Robert; Kuo, Alan; Andreopoulos, William; LaButti, Kurt; Pangilinan, Jasmyn; Tritt, Andrew; Lipzen, Anna; He, Guifen; Yan, Mi; Vivian, Ng; Grigoriev, Igor V.; Cullen, Daniel; Martin, Francis; Rosso, Marie-Noëlle; Henrissat, Bernard; Hibbett, David; Martínez, Ángel T.; Institute for Multidisciplinary Research in Applied Biology - IMAB
As actors of global carbon cycle, Agaricomycetes (Basidiomycota) have developed complex enzymatic machineries that allow them to decompose all plant polymers, including lignin. Among them, saprotrophic Agaricales are characterized by an unparalleled diversity of habitats and lifestyles. Comparative analysis of 52 Agaricomycetes genomes (14 of them sequenced de novo) reveals that Agaricales possess a large diversity of hydrolytic and oxidative enzymes for lignocellulose decay. Based on the gene families with the predicted highest evolutionary rates-namely cellulose-binding CBM1, glycoside hydrolase GH43, lytic polysaccharide monooxygenase AA9, class-II peroxidases, glucose-methanol-choline oxidase/dehydrogenases, laccases, and unspecific peroxygenases-we reconstructed the lifestyles of the ancestors that led to the extant lignocellulose-decomposing Agaricomycetes. The changes in the enzymatic toolkit of ancestral Agaricales are correlated with the evolution of their ability to grow not only on wood but also on leaf litter and decayed wood, with grass-litter decomposers as the most recent eco-physiological group. In this context, the above families were analyzed in detail in connection with lifestyle diversity. Peroxidases appear as a central component of the enzymatic toolkit of saprotrophic Agaricomycetes, consistent with their essential role in lignin degradation and high evolutionary rates. This includes not only expansions/losses in peroxidase genes common to other basidiomycetes but also the widespread presence in Agaricales (and Russulales) of new peroxidases types not found in wood-rotting Polyporales, and other Agaricomycetes orders. Therefore, we analyzed the peroxidase evolution in Agaricomycetes by ancestralsequence reconstruction revealing several major evolutionary pathways and mapped the appearance of the different enzyme types in a time-calibrated species tree.
Open Access
A secretomic view of woody and nonwoody lignocellulose degradation by Pleurotus ostreatus
(BioMed Central, 2016) Fernández Fueyo, Elena; Ruiz Dueñas, Francisco J.; López Lucendo, María F.; Pérez Boada, Marta; Rencoret, Jorge; Gutiérrez, Ana; Pisabarro de Lucas, Gerardo; Ramírez Nasto, Lucía; Martínez, Ángel T.; Producción Agraria; Nekazaritza Ekoizpena
Background: Pleurotus ostreatus is the second edible mushroom worldwide, and a model fungus for delignification applications, with the advantage of growing on woody and nonwoody feedstocks. Its sequenced genome is available, and this gave us the opportunity to perform proteomic studies to identify the enzymes overproduced in lignocellulose cultures. Results: Monokaryotic P. ostreatus (PC9) was grown with poplar wood or wheat straw as the sole C/N source and the extracellular proteins were analyzed, together with those from glucose medium. Using nano-liquid chromatography coupled to tandem mass spectrometry of whole-protein hydrolyzate, over five-hundred proteins were identified. Thirty-four percent were unique of the straw cultures, while only 15 and 6 % were unique of the glucose and poplar cultures, respectively (20 % were produced under the three conditions, and additional 19 % were shared by the two lignocellulose cultures). Semi-quantitative analysis showed oxidoreductases as the main protein type both in the poplar (39 % total abundance) and straw (31 %) secretomes, while carbohydrate-active enzymes (CAZys) were only slightly overproduced (14–16 %). Laccase 10 (LACC10) was the main protein in the two lignocellulose secretomes (10–14 %) and, together with LACC2, LACC9, LACC6, versatile peroxidase 1 (VP1), and manganese peroxidase 3 (MnP3), were strongly overproduced in the lignocellulose cultures. Seven CAZys were also among the top-50 proteins, but only CE16 acetylesterase was overproduced on lignocellulose. When the woody and nonwoody secretomes were compared, GH1 and GH3 β-glycosidases were more abundant on poplar and straw, respectively and, among less abundant proteins, VP2 was overproduced on straw, while VP3 was only found on poplar. The treated lignocellulosic substrates were analyzed by two-dimensional nuclear magnetic resonance (2D NMR), and a decrease of lignin relative to carbohydrate signals was observed, together with the disappearance of some minor lignin substructures, and an increase of sugar reducing ends. Conclusions: Oxidoreductases are strongly induced when P. ostreatus grows on woody and nonwoody lignocellulosic substrates. One laccase occupied the first position in both secretomes, and three more were overproduced together with one VP and one MnP, suggesting an important role in lignocellulose degradation. Preferential removal of lignin vs carbohydrates was shown by 2D NMR, in agreement with the above secretomic results.

Ramírez Nasto, Lucía

Email Address

person.page.identifierURI

Birth Date

Job Title

Last Name

First Name

person.page.departamento

person.page.instituteName

ORCID

person.page.observainves

person.page.upna

Name

Filters

Author

Subject

Date

Has files

Item Type

Type

Settings

Sort By

Results per page

Search Results