Mapping N-linked Glycosylation Of Carbohydrate-active Enzymes In The Secretome Of Aspergillus Nidulans Grown On Lignocellulose

dc.creatorRubio
dc.creatorMarcelo Ventura; Zubieta
dc.creatorMariane Paludetti; Loureno Franco Cairo
dc.creatorJoao Paulo; Calzado
dc.creatorFelipe; Paes Leme
dc.creatorAdriana Franco; Squina
dc.creatorFabio Marcio; Prade
dc.creatorRolf Alexander; de Lima Damasio
dc.creatorAndre Ricardo
dc.date2016
dc.dateagos
dc.date2017-11-13T13:24:33Z
dc.date2017-11-13T13:24:33Z
dc.date.accessioned2018-03-29T05:57:04Z
dc.date.available2018-03-29T05:57:04Z
dc.identifierBiotechnology For Biofuels. Biomed Central Ltd, v. 9, p. , 2016.
dc.identifier1754-6834
dc.identifierWOS:000380876500001
dc.identifier10.1186/s13068-016-0580-4
dc.identifierhttps://biotechnologyforbiofuels.biomedcentral.com.ez88.periodicos.capes.gov.br/articles/10.1186/s13068-016-0580-4
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/328323
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1365348
dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.descriptionConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.descriptionThe genus Aspergillus includes microorganisms that naturally degrade lignocellulosic biomass, secreting large amounts of carbohydrate-active enzymes (CAZymes) that characterize their saprophyte lifestyle. Aspergillus has the capacity to perform post-translational modifications (PTM), which provides an additional advantage for the use of these organisms as a host for the production of heterologous proteins. In this study, the N-linked glycosylation of CAZymes identified in the secretome of Aspergillus nidulans grown on lignocellulose was mapped. Results: Aspergillus nidulans was grown in glucose, xylan and pretreated sugarcane bagasse (SCB) for 96 h, after which glycoproteomics and glycomics were carried out on the extracellular proteins (secretome). A total of 265 proteins were identified, with 153, 210 and 182 proteins in the glucose, xylan and SCB substrates, respectively. CAZymes corresponded to more than 50 % of the total secretome in xylan and SCB. A total of 182 N-glycosylation sites were identified, of which 121 were detected in 67 CAZymes. A prevalence of the N-glyc sequon N-X-T (72.2 %) was observed in N-glyc sites compared with N-X-S (27.8 %). The amino acids flanking the validated N-glyc sites were mainly composed of hydrophobic and polar uncharged amino acids. Selected proteins were evaluated for conservation of the N-glyc sites in Aspergilli homologous proteins, but a pattern of conservation was not observed. A global analysis of N-glycans released from the proteins secreted by A. nidulans was also performed. While the proportion of N-glycans with Hex5 to Hex9 was similar in the xylan condition, a prevalence of Hex5 was observed in the SCB and glucose conditions. Conclusions: The most common and frequent N-glycosylated motifs, an overview of the N-glycosylation of the CAZymes and the number of mannoses found in N-glycans were analyzed. There are many bottlenecks in protein production by filamentous fungi, such as folding, transport by vesicles and secretion, but N-glycosylation in the correct context is a fundamental event for defining the high levels of secretion of target proteins. A comprehensive analysis of the protein glycosylation processes in A. nidulans will assist with a better understanding of glycoprotein structures, profiles, activities and functions. This knowledge can help in the optimization of heterologous expression and protein secretion in the fungal host.
dc.description9
dc.descriptionFAPESP [2012/20549-4, 2014/06923-6, 2009/54067-3, 2013/24988-5, 2014/15403-6, 2011/20977-3, 2014/23051-2]
dc.descriptionNational Institutes of Health (NIH/NCRR) [P41GM10349010]
dc.descriptionNational Council for Scientific and Technological Development (CNPq) [441912/2014-1, 310186/2014-5, 442333/2014-5]
dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.descriptionConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.languageEnglish
dc.publisherBiomed Central Ltd
dc.publisherLondon
dc.relationBiotechnology for Biofuels
dc.rightsfechado
dc.sourceWOS
dc.subjectGlycoproteomics
dc.subjectAspergillus Nidulans
dc.subjectCarbohydrate-active Enzymes
dc.subjectCazy
dc.subjectGlycoside Hydrolases
dc.subjectN-glycosylation
dc.subjectHeterologous Expression
dc.titleMapping N-linked Glycosylation Of Carbohydrate-active Enzymes In The Secretome Of Aspergillus Nidulans Grown On Lignocellulose
dc.typeArtículos de revistas


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