Structure, Dynamics, and Specificity of Endoglucanase D from Clostridium cellulovorans

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dc.citation.firstpage4267
dc.citation.issueNumber22
dc.citation.journalTitleJournal of Molecular Biology
dc.citation.lastpage4285
dc.citation.volumeNumber425
dc.contributor.authorBianchetti, Christopher M.
dc.contributor.authorBrumm, Phillip
dc.contributor.authorSmith, Robert W.
dc.contributor.authorDyer, Kevin
dc.contributor.authorHura, Greg L.
dc.contributor.authorRutkoski, Thomas J.
dc.contributor.authorPhillips, George N.Jr.
dc.date.accessioned2017-08-04T12:30:01Z
dc.date.available2017-08-04T12:30:01Z
dc.date.issued2013
dc.description.abstractThe enzymatic degradation of cellulose is a critical step in the biological conversion of plant biomass into an abundant renewable energy source. An understanding of the structural and dynamic features that cellulases utilize to bind a single strand of crystalline cellulose and hydrolyze the β-1,4-glycosidic bonds of cellulose to produce fermentable sugars would greatly facilitate the engineering of improved cellulases for the large-scale conversion of plant biomass. Endoglucanase D (EngD) from Clostridium cellulovorans is a modular enzyme comprising an N-terminal catalytic domain and a C-terminal carbohydrate-binding module, which is attached via a flexible linker. Here, we present the 2.1-Å-resolution crystal structures of full-length EngD with and without cellotriose bound, solution small-angle X-ray scattering (SAXS) studies of the full-length enzyme, the characterization of the active cleft glucose binding subsites, and substrate specificity of EngD on soluble and insoluble polymeric carbohydrates. SAXS data support a model in which the linker is flexible, allowing EngD to adopt an extended conformation in solution. The cellotriose-bound EngD structure revealed an extended active-site cleft that contains seven glucose-binding subsites, but unlike the majority of structurally determined endocellulases, the active-site cleft of EngD is partially enclosed by Trp162 and Tyr232. EngD variants, which lack Trp162, showed a significant reduction in activity and an alteration in the distribution of cellohexaose degradation products, suggesting that Trp162 plays a direct role in substrate binding.
dc.identifier.citationBianchetti, Christopher M., Brumm, Phillip, Smith, Robert W., et al.. "Structure, Dynamics, and Specificity of Endoglucanase D from Clostridium cellulovorans." <i>Journal of Molecular Biology,</i> 425, no. 22 (2013) Elsevier: 4267-4285. https://doi.org/10.1016/j.jmb.2013.05.030.
dc.identifier.digitalStructure_Dynamics_Specificity
dc.identifier.doihttps://doi.org/10.1016/j.jmb.2013.05.030
dc.identifier.urihttps://hdl.handle.net/1911/96591
dc.language.isoeng
dc.publisherElsevier
dc.rightsThis is an author's peer-reviewed final manuscript, as accepted by the publisher. The published article is copyrighted by Elsevier.
dc.subject.keywordCBM
dc.subject.keywordEngD
dc.subject.keywordGH
dc.subject.keywordPDB
dc.subject.keywordPT
dc.subject.keywordProtein Data Bank
dc.subject.keywordSAXS
dc.subject.keywordX-ray crystallography
dc.subject.keywordcarbohydrate-binding module
dc.subject.keywordcellulase
dc.subject.keywordcellulose degradation
dc.subject.keywordendoglucanase
dc.subject.keywordendoglucanase D
dc.subject.keywordglycosyl hydrolase
dc.subject.keywordproline/threonine-rich
dc.subject.keywordsmall-angle X-ray scattering
dc.titleStructure, Dynamics, and Specificity of Endoglucanase D from Clostridium cellulovorans
dc.typeJournal article
dc.type.dcmiText
dc.type.publicationpost-print
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