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Characterization of Multifunctional Enzyme Complexes from Comamonas testosteroni KF1 and Thermomonospora curvata DSM 43183 Involved in Steroid Side Chain Degradation

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dc.contributor.advisor Seah, Stephen
dc.contributor.author Aggett, Rebecca
dc.date.accessioned 2018-12-17T17:38:08Z
dc.date.copyright 2018-11
dc.date.created 2018-12-05
dc.date.issued 2018-12-17
dc.identifier.uri http://hdl.handle.net/10214/14655
dc.description.abstract The genes fadI and fadJ from Comamonas testosteroni KF1 are homologous to genes encoding the components of multifunctional enzyme complexes involved in fatty acid side chain β-oxidation. Both of these genes are located within the steroid degradation locus in the genome. FadJ had hydratase activity towards the 5-carbon side chain of cholic acid metabolite; however, gene knockout studies revealed no phenotypic changes in growth patterns. In the Actinobacterium Thermomonospora curvata DSM 43183, the genes Tcur3480, Tcur3482 and ltp2Tcur, located within a steroid side chain degrading operon, together encode a multifunctional steroid degrading enzyme complex. It was previously determined that the aldolase Ltp2Tcur associates with the hydratase Tcur3480-Tcur3482 through the DUF35 domain, although the catalytic mechanism and kinetic parameters were unknown. The crystal structure of Ltp2Tcur-DUF35 was solved and wild-type and mutant variant kinetic parameters were determined to support a proposed, novel retro-aldol catalytic mechanism involving two catalytic tyrosine residues. en_US
dc.description.sponsorship Research supported by grant 2015-05366 from the Natural Science and Engineering Research Council of Canada. en_US
dc.language.iso en en_US
dc.subject site specific mutagenesis en_US
dc.subject gene knockout en_US
dc.subject steroid en_US
dc.subject aldolase en_US
dc.subject hydratase en_US
dc.subject crystallography en_US
dc.title Characterization of Multifunctional Enzyme Complexes from Comamonas testosteroni KF1 and Thermomonospora curvata DSM 43183 Involved in Steroid Side Chain Degradation en_US
dc.type Thesis en_US
dc.degree.programme Molecular and Cellular Biology en_US
dc.degree.name Master of Science en_US
dc.degree.department Department of Molecular and Cellular Biology en_US
dc.description.embargo 2019-12-05


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