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dc.contributor.authorChandrayan, Puja
dc.date.accessioned2014-05-06T04:30:16Z
dc.date.available2014-05-06T04:30:16Z
dc.date.issued2013-12
dc.identifier.otherchandrayan_puja_201312_ms
dc.identifier.urihttp://purl.galileo.usg.edu/uga_etd/chandrayan_puja_201312_ms
dc.identifier.urihttp://hdl.handle.net/10724/29729
dc.description.abstractCaldicellulosiruptor bescii is a gram-positive thermophilic bacterium with the ability to grow on untreated plant biomass. Upon growth on switchgrass at 78°C, a set of genes encoding 94 proteins was up-regulated transcriptionally by more than 4-fold, in comparison to growth on glucose. Enzymes with activity towards pectin were among those that were most highly up-regulated. Of a total of four predicted pectinolytic enzymes, Cbes_1854, a multidomain protein of polysaccharide lyase family 3, was chosen for overexpression and purification in E. coli. The full-length protein (X-PL3) and just the catalytic domain (PL3) were purified with yields of approximately 10 mg/liter E. coli culture. Both purified recombinant proteins display alkaline pH optima and have a half-life of approximately one hour at 90°C. Under in vitro assay conditions, domain X does not play a significant role in catalysis. The catalytic domain, PL3, also shows a synergistic effect on sugar release from untreated switchgrass biomass during in vitro assays with a cellulase (Cel A) from C. bescii. In a collaborative study, the crystal structure of the PL3 domain was obtained. This provided insight into the catalytic mechanism of the enzyme.
dc.languageeng
dc.publisheruga
dc.rightspublic
dc.subjectCaldicellulosiruptor bescii, Cbes_1854
dc.subjectPectinolytic enzymes, Polysaccharide lyase family 3
dc.subjectCatalytic domain (PL3), Full-length protein (X-PL3)
dc.subjectE. coli, Cellulase (Cel A)
dc.titleBiochemical characterization of a pectate lyase from Caldicellulosiruptor bescii
dc.typeThesis
dc.description.degreeMS
dc.description.departmentBiochemistry and Molecular Biology
dc.description.majorBiochemistry and Molecular Biology
dc.description.advisorMichael W.w. Adams
dc.description.committeeMichael W.w. Adams
dc.description.committeeRobert Maier
dc.description.committeeWilliam Lanzilotta


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