Understanding the Function of Conserved Variations in the Catalytic Loops of Fungal Glycoside Hydrolase Family 12

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dc.contributor.authorDamasio, Andre R. L.
dc.contributor.authorRubio, Marcelo V.
dc.contributor.authorOliveira, Leandro C. [UNESP]
dc.contributor.authorSegato, Fernando
dc.contributor.authorDias, Bruno A. [UNESP]
dc.contributor.authorCitadini, Ana P.
dc.contributor.authorPaixao, Douglas A.
dc.contributor.authorSquina, Fabio M.
dc.contributor.institutionCtr Nacl Pesquisa Energia & Mat CNPEM
dc.contributor.institutionUniversidade Estadual de Campinas (UNICAMP)
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.date.accessioned2015-03-18T15:52:32Z
dc.date.available2015-03-18T15:52:32Z
dc.date.issued2014-08-01
dc.description.abstractEnzymes that cleave the xyloglucan backbone at unbranched glucose residues have been identified in GH families 5, 7, 12, 16, 44, and 74. Fungi produce enzymes that populate 20 of 22 families that are considered critical for plant biomass deconstruction. We searched for GH12-encoding genes in 27 Eurotiomycetes genomes. After analyzing 50 GH12-related sequences, the conserved variations of the amino acid sequences were examined. Compared to the endoglucanases, the endo-xyloglucanase-associated YSG deletion at the negative subsites of the catalytic cleft with a SST insertion at the reducing end of the substrate-binding crevice is highly conserved. In addition, a highly conserved alanine residue was identified in all xyloglucan-specific enzymes, and this residue is substituted by arginine in more promiscuous glucanases. To understand the basis for the xyloglucan specificity displayed by certain GH12 enzymes, two fungal GH12 endoglucanases were chosen for mutagenesis and functional studies: an endo-xyloglucanase from Aspergillus clavatus (AclaXegA) and an endoglucanase from A. terreus (AtEglD). Comprehensive molecular docking studies and biochemical analyses were performed, revealing that mutations at the entrance of the catalytic cleft in AtEglD result in a wider binding cleft and the alteration of the substrate-cleavage pattern, implying that a trio of residues coordinates the interactions and binding to linear glycans. The loop insertion at the crevice-reducing end of AclaXegA is critical for catalytic efficiency to hydrolyze xyloglucan. The understanding of the structural elements governing endo-xyloglucanase activity on linear and branched glucans will facilitate future enzyme modifications with potential applications in industrial biotechnology. (C) 2014 Wiley Periodicals, Inc.en
dc.description.affiliationCtr Nacl Pesquisa Energia & Mat CNPEM, Lab Nacl Ciencia Tecnol Bioetanol CTBE, Campinas, SP, Brazil
dc.description.affiliationUniv Estadual Campinas UNICAMP, Inst Biol, Campinas, SP, Brazil
dc.description.affiliationUniv Estadual Paulista UNESP, Inst Biociencias Letras & Ciencias Exatas, Sao Jose Do Rio Preto, SP, Brazil
dc.description.affiliationUnespUniv Estadual Paulista UNESP, Inst Biociencias Letras & Ciencias Exatas, Sao Jose Do Rio Preto, SP, Brazil
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipIdCNPq: 474022/2011-4
dc.description.sponsorshipIdCNPq: 310177/2011-1
dc.description.sponsorshipIdFAPESP: 08/58037-9
dc.description.sponsorshipIdFAPESP: 11/02169-4
dc.description.sponsorshipIdFAPESP: 11/13242-7
dc.description.sponsorshipIdFAPESP: 12/12859-3
dc.format.extent1494-1505
dc.identifierhttp://dx.doi.org/10.1002/bit.25209
dc.identifier.citationBiotechnology And Bioengineering. Hoboken: Wiley-blackwell, v. 111, n. 8, p. 1494-1505, 2014.
dc.identifier.doi10.1002/bit.25209
dc.identifier.issn0006-3592
dc.identifier.urihttp://hdl.handle.net/11449/116178
dc.identifier.wosWOS:000341235400003
dc.language.isoeng
dc.publisherWiley-Blackwell
dc.relation.ispartofBiotechnology And Bioengineering
dc.relation.ispartofjcr3.952
dc.relation.ispartofsjr1,372
dc.rights.accessRightsAcesso restrito
dc.sourceWeb of Science
dc.subjectfungal endoglucanasesen
dc.subjectGH12en
dc.subjectendo-xyloglucanasesen
dc.subjectxyloglucan specificityen
dc.titleUnderstanding the Function of Conserved Variations in the Catalytic Loops of Fungal Glycoside Hydrolase Family 12en
dc.typeArtigo
dcterms.licensehttp://olabout.wiley.com/WileyCDA/Section/id-406071.html
dcterms.rightsHolderWiley-Blackwell

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