Structure and function of enzymes and auxiliary proteins from Trichoderma, active in cell-wall hydrolysis

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dc.contributorInstituto de Física de São Carlos – IFSC/USPpt_BR
dc.contributor.authorPolikarpov, Igor
dc.date.accessioned2017-05-29T13:09:56Z
dc.date.available2017-05-29T13:09:56Z
dc.date.issued2017-05-29
dc.description.abstractLignocellulosic biomass, such as sugarcane bagasse, holds a promise of environmentally friendly bioenergy production in Brazil. However, enzymatic hydrolysis, currently considered a method of choice in biomass saccharification, is hampered by considerable cell-wall recalcitrance. To make this technology sustainable and cost effective, our comprehension of cellulose enzymatic hydrolysis should be significantly improved. Here we propose to conduct systematic structure-functional studies of Trichoderma cellulases and auxiliary proteins active in cell-wall degradation using a combination of X-ray protein crystallography, biophysical and biochemical studies, molecular dynamics simulations, statistical coupling analysis aligned with the site-directed mutagenesis and enzymatic assays aiming to obtain in-depth comprehension of cellulose hydrolysis. We plan to contribute toward structural analysis of Trichoderma reesei endoglucanases by solving a crystal structure of endoglucanase II (Cel5A), main enzymatically active, but structurally uncharacterized endoglucanase of this important industrial fungus. Moreover, we will contribute toward our knowledge of Trichoderma cellulases molecular organization by solving X-ray structures of main Trichoderma harzianum endo- and exoglucanases (primarily focusing on Cel7A and Cel5A) and by comparing them with the correspondent T. reesei enzymes. We also aim to structurally characterize swollenins, non-hydrolytic proteins, shown to enhance cellulose hydrolysis catalyzed by celulases, and to study thermodynamically its interactions with cellulose. In addition, we will construct chimeric enzymes by fusing of swollenin with the cellulases and will study enzymatic properties of such chimeras. Furthermore, we will conduct systematic molecular dynamics studies of the cellulases and swollenin, and investigate their flexibility by hydrogen deuterium exchange followed by massspectrometry. Finally, we will use all these acquired knowledge to modify the proteins using site-directed mutagenesis aiming to better comprehend molecular basis of their function and to produce enzymes and their mixtures with enhanced hydrolytic properties.pt_BR
dc.description.notesOutorgado: Prof. Dr. Igor Polikarpov; Universidade de São Paulo, USP, Instituto de Física de São Carlos, IFSC, Departamento de Física e Ciência Interdisciplinar, FCI, Grupo de Biotecnologia Molecular, São Carlos, SP, Brasil.pt_BR
dc.description.sponsorshipFAPESP (08/56255-9)pt_BR
dc.format3 p.pt_BR
dc.format.mediumDigitalpt_BR
dc.identifier.urihttp://repositorio.ifsc.usp.br/handle/RIIFSC/8921
dc.language.isoengpt_BR
dc.rightsAcesso abertopt_BR
dc.subjectBiologia molecularpt_BR
dc.subjectFungos mitospóricospt_BR
dc.subjectTrichodermapt_BR
dc.subjectEnzimaspt_BR
dc.subject.classificationIFSC - FCIpt_BR
dc.titleStructure and function of enzymes and auxiliary proteins from Trichoderma, active in cell-wall hydrolysispt_BR
dc.type.categoryPesquisapt_BR
usp.date.end2014-09-30
usp.date.initial2009-11-01
usp.date.ratification2009
usp.description.localSão Carlos, SP, Brasilpt_BR
usp.isreferencedbyhttp://www.bv.fapesp.br/pt/auxilios/6479/structure-and-function-of-enzymes-and-auxiliary-proteins-from-trichoderma-active-in-cell-wall-hydro/pt_BR
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