A secretomic view of woody and nonwoody lignocellulose degradation by Pleurotus ostreatus

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.

A secretomic view of woody and nonwoody lignocellulose degradation by Pleurotus ostreatus

dc.contributor.author	Fernández Fueyo, Elena
dc.contributor.author	Ruiz Dueñas, Francisco J.
dc.contributor.author	López Lucendo, María F.
dc.contributor.author	Pérez Boada, Marta
dc.contributor.author	Rencoret, Jorge
dc.contributor.author	Gutiérrez, Ana
dc.contributor.author	Pisabarro de Lucas, Gerardo
dc.contributor.author	Ramírez Nasto, Lucía
dc.contributor.author	Martínez, Ángel T.
dc.contributor.department	Producción Agraria	es_ES
dc.contributor.department	Nekazaritza Ekoizpena	eu
dc.date.accessioned	2018-08-31T08:07:58Z
dc.date.available	2018-08-31T08:07:58Z
dc.date.issued	2016
dc.description.abstract	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.	en
dc.description.sponsorship	This work was supported by the INDOX (KBBE-2013-613549) EU project, the BIO2014-56388-R, AGL2014-53730-R, and AGL2011-30495 projects of the Spanish Ministry of Economy and Competitiveness (MINECO) co-financed by FEDER funds, and the ProteoRed platform of the Spanish Institute of Health Carlos III (ISCIII). The work conducted by the US DOE JGI is supported by the Office of Science of the US DOE under contract number DE-AC02-05CH11231. FJR-D thanks a Ramón y Cajal contract of the Spanish MINECO, and JR thanks a contract of the CSIC project 201440E097.	en
dc.format.extent	18 p.
dc.format.mimetype	application/pdf	en
dc.format.mimetype	application/zip	en
dc.identifier.doi	10.1186/s13068-016-0462-9
dc.identifier.issn	1754-6834
dc.identifier.uri	https://academica-e.unavarra.es/handle/2454/30350
dc.language.iso	eng	en
dc.publisher	BioMed Central	en
dc.relation.ispartof	Biotechnology for Biofuels, (2016) 9:49	en
dc.relation.projectID	info:eu-repo/grantAgreement/MINECO//BIO2014-56388-R/ES/
dc.relation.projectID	info:eu-repo/grantAgreement/MINECO//AGL2014-53730-R/ES/
dc.relation.projectID	info:eu-repo/grantAgreement/MICINN//AGL2011-30495/ES/
dc.relation.projectID	info:eu-repo/grantAgreement/European Commission/FP7/613549/
dc.relation.publisherversion	https://doi.org/10.1186/s13068-016-0462-9
dc.rights	© 2016 Fernández-Fueyo et al. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons. org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.	en
dc.rights.accessRights	info:eu-repo/semantics/openAccess
dc.rights.uri	https://creativecommons.org/licenses/by/4.0/
dc.subject	Pleurotus ostreatus	en
dc.subject	Secreted proteins	en
dc.subject	Poplar wood	en
dc.subject	Wheat straw	en
dc.subject	LC–MS/MS	en
dc.subject	Lignin-modifying enzymes	en
dc.subject	Laccases	en
dc.subject	Carbohydrate-active enzymes	en
dc.subject	2D NMR	en
dc.title	A secretomic view of woody and nonwoody lignocellulose degradation by Pleurotus ostreatus	en
dc.type	info:eu-repo/semantics/article
dc.type.version	info:eu-repo/semantics/publishedVersion
dspace.entity.type	Publication
relation.isAuthorOfPublication	54daf1c4-f273-4ade-a90e-7df28e761ca7
relation.isAuthorOfPublication	7fedbadd-7e13-485e-99cf-252f1a381373
relation.isAuthorOfPublication.latestForDiscovery	54daf1c4-f273-4ade-a90e-7df28e761ca7