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dc.contributor.authorBuddrus, Lisaen_NZ
dc.contributor.authorAndrews, Emma Sophie Vouten_NZ
dc.contributor.authorLeak, David J.en_NZ
dc.contributor.authorDanson, Michael J.en_NZ
dc.contributor.authorArcus, Vickery L.en_NZ
dc.contributor.authorCrennell, Susan J.en_NZ
dc.date.accessioned2018-06-07T00:08:56Z
dc.date.available2018-03-01en_NZ
dc.date.available2018-06-07T00:08:56Z
dc.date.issued2018en_NZ
dc.identifier.citationBuddrus, L., Andrews, E. S. V., Leak, D. J., Danson, M. J., Arcus, V. L., & Crennell, S. J. (2018). Crystal structure of an inferred ancestral bacterial pyruvate decarboxylase. Acta Crystallographica Section F: Structural Biology Communications, 74, 179–186. https://doi.org/10.1107/S2053230X18002819en
dc.identifier.issn2053-230Xen_NZ
dc.identifier.urihttps://hdl.handle.net/10289/11883
dc.description.abstractPyruvate decarboxylase (PDC; EC 4.1.1.1) is a key enzyme in homofermentative metabolism where ethanol is the major product. PDCs are thiamine pyrophos­phate- and Mg2+ ion-dependent enzymes that catalyse the non-oxidative decarboxylation of pyruvate to acetaldehyde and carbon dioxide. As this enzyme class is rare in bacteria, current knowledge of bacterial PDCs is extremely limited. One approach to further the understanding of bacterial PDCs is to exploit the diversity provided by evolution. Ancestral sequence reconstruction (ASR) is a method of computational molecular evolution to infer extinct ancestral protein sequences, which can then be synthesized and experimentally characterized. Through ASR a novel PDC was generated, designated ANC27, that shares only 78% amino-acid sequence identity with its closest extant homologue (Komagataeibacter medellinensis PDC, GenBank accession No. WP_014105323.1), yet is fully functional. Crystals of this PDC diffracted to 3.5 Å resolution. The data were merged in space group P3221, with unit-cell parameters a = b = 108.33, c = 322.65 Å, and contained two dimers (two tetramer halves) in the asymmetric unit. The structure was solved by molecular replacement using PDB entry 2wvg as a model, and the final R values were Rwork = 0.246 (0.3671 in the highest resolution bin) and Rfree = 0.319 (0.4482 in the highest resolution bin). Comparison with extant bacterial PDCs supports the previously observed correlation between decreased tetramer interface area (and number of interactions) and decreased thermostability.
dc.format.mimetypeapplication/pdf
dc.language.isoenen_NZ
dc.publisherInternational Union of Crystallographyen_NZ
dc.rightsThis is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.
dc.subjectScience & Technologyen_NZ
dc.subjectLife Sciences & Biomedicineen_NZ
dc.subjectPhysical Sciencesen_NZ
dc.subjectBiochemical Research Methodsen_NZ
dc.subjectBiochemistry & Molecular Biologyen_NZ
dc.subjectBiophysicsen_NZ
dc.subjectCrystallographyen_NZ
dc.subjectancestral sequence reconstructionen_NZ
dc.subjectpyruvate decarboxylaseen_NZ
dc.subjectlyasesen_NZ
dc.subjectcrystal structureen_NZ
dc.subjectTPP-dependent enzymesen_NZ
dc.subjectZYMOMONAS-MOBILISen_NZ
dc.subjectPHYLOGENETIC ANALYSISen_NZ
dc.subjectMAXIMUM-LIKELIHOODen_NZ
dc.subjectZYMOBACTER-PALMAEen_NZ
dc.subjectEVOLUTIONen_NZ
dc.subjectENZYMESen_NZ
dc.subjectLIFEen_NZ
dc.subjectRECONSTRUCTIONen_NZ
dc.subjectRESOLUTIONen_NZ
dc.subjectINTERFACEen_NZ
dc.titleCrystal structure of an inferred ancestral bacterial pyruvate decarboxylaseen_NZ
dc.typeJournal Article
dc.identifier.doi10.1107/S2053230X18002819en_NZ
dc.relation.isPartOfActa Crystallographica Section F: Structural Biology Communicationsen_NZ
pubs.begin-page179
pubs.elements-id220230
pubs.end-page186
pubs.publication-statusPublisheden_NZ
pubs.volume74en_NZ


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