A ‘Split-Gene’ Transketolase From the Hyper-Thermophilic Bacterium Carboxydothermus hydrogenoformans: Structure and Biochemical Characterization
dc.contributor.author | James, P | |
dc.contributor.author | Isupov, MN | |
dc.contributor.author | De Rose, SA | |
dc.contributor.author | Sayer, C | |
dc.contributor.author | Cole, IS | |
dc.contributor.author | Littlechild, JA | |
dc.date.accessioned | 2020-11-09T08:53:32Z | |
dc.date.issued | 2020-10-30 | |
dc.description.abstract | A novel transketolase has been reconstituted from two separate polypeptide chains encoded by a ‘split-gene’ identified in the genome of the hyperthermophilic bacterium, Carboxydothermus hydrogenoformans. The reconstituted active α2β2 tetrameric enzyme has been biochemically characterized and its activity has been determined using a range of aldehydes including glycolaldehyde, phenylacetaldehyde and cyclohexanecarboxaldehyde as the ketol acceptor and hydroxypyruvate as the donor. This reaction proceeds to near 100% completion due to the release of the product carbon dioxide and can be used for the synthesis of a range of sugars of interest to the pharmaceutical industry. This novel reconstituted transketolase is thermally stable with no loss of activity after incubation for 1 h at 70°C and is stable after 1 h incubation with 50% of the organic solvents methanol, ethanol, isopropanol, DMSO, acetonitrile and acetone. The X-ray structure of the holo reconstituted α2β2 tetrameric transketolase has been determined to 1.4 Å resolution. In addition, the structure of an inactive tetrameric β4 protein has been determined to 1.9 Å resolution. The structure of the active reconstituted α2β2 enzyme has been compared to the structures of related enzymes; the E1 component of the pyruvate dehydrogenase complex and D-xylulose-5-phosphate synthase, in an attempt to rationalize differences in structure and substrate specificity between these enzymes. This is the first example of a reconstituted ‘split-gene’ transketolase to be biochemically and structurally characterized allowing its potential for industrial biocatalysis to be evaluated. | en_GB |
dc.description.sponsorship | Biotechnology & Biological Sciences Research Council (BBSRC) | en_GB |
dc.identifier.citation | Vol. 11, article 592353 | en_GB |
dc.identifier.doi | 10.3389/fmicb.2020.592353 | |
dc.identifier.grantnumber | BB/L002035/1 | en_GB |
dc.identifier.uri | http://hdl.handle.net/10871/123528 | |
dc.language.iso | en | en_GB |
dc.publisher | Frontiers Media | en_GB |
dc.relation.url | http://www.wwpdb.org/ | en_GB |
dc.rights | © 2020 James, Isupov, De Rose, Sayer, Cole and Littlechild. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. | en_GB |
dc.subject | hyperthermophilic | en_GB |
dc.subject | ‘split-gene’ | en_GB |
dc.subject | transketolase | en_GB |
dc.subject | thermal stability | en_GB |
dc.subject | industrial applications | en_GB |
dc.title | A ‘Split-Gene’ Transketolase From the Hyper-Thermophilic Bacterium Carboxydothermus hydrogenoformans: Structure and Biochemical Characterization | en_GB |
dc.type | Article | en_GB |
dc.date.available | 2020-11-09T08:53:32Z | |
dc.description | This is the final version. Available on open access from Frontiers Media via the DOI in this record | en_GB |
dc.description | Data Availability Statement: The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found below: http://www.wwpdb.org/, 6YAK and 6YAJ | en_GB |
dc.identifier.eissn | 1664-302X | |
dc.identifier.journal | Frontiers in Microbiology | en_GB |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0/ | en_GB |
dcterms.dateAccepted | 2020-10-12 | |
exeter.funder | ::Biotechnology & Biological Sciences Research Council (BBSRC) | en_GB |
rioxxterms.version | VoR | en_GB |
rioxxterms.licenseref.startdate | 2020-10-12 | |
rioxxterms.type | Journal Article/Review | en_GB |
refterms.dateFCD | 2020-11-09T08:52:05Z | |
refterms.versionFCD | VoR | |
refterms.dateFOA | 2020-11-09T08:53:35Z | |
refterms.panel | A | en_GB |
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Except where otherwise noted, this item's licence is described as © 2020 James, Isupov, De Rose, Sayer, Cole and Littlechild. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.