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http://hdl.handle.net/11375/21158
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DC Field | Value | Language |
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dc.contributor.advisor | Brown, Eric | - |
dc.contributor.author | Allison, Sarah | - |
dc.date.accessioned | 2017-03-02T16:57:25Z | - |
dc.date.available | 2017-03-02T16:57:25Z | - |
dc.date.issued | 2011-04 | - |
dc.identifier.uri | http://hdl.handle.net/11375/21158 | - |
dc.description.abstract | The biosynthetic enzymes involved in wall teichoic acid biogenesis in Grampositive bacteria have been the subject of renewed investigation in recent years with the benefit of modem tools of biochemistry and genetics. Nevertheless, there have been only limited investigations into the enzymes that glycosylate wall teichoic acid. Decades-old experiments in the model Gram-positive bacterium, Bacillus subtilis 168, using phage resistant mutants implicated tagE (also called gtaA and rodD) as the gene coding for the wall teichoic acid glycosyltransferase. This study and others have provided only indirect evidence to support a role for TagE in wall teichoic acid glycosylation. In this work, we showed that deletion of tagE results in the loss of a-glucose at the C-2 position of glycerol in the poly(glycerol phosphate) polymer backbone. We also report the first kinetic characterization of pure, recombinant wall teichoic acid glycosyltransferase using clean synthetic substrates. We investigated the substrate specificity ofTagE using a wide variety of acceptor substrates and showed that this enzyme has a strong kinetic preference for the transfer of glucose from UDP-glucose to glycerol phosphate in polymeric form. Further, we showed that the enzyme recognizes its polymeric (and repetitive) substrate with a sequential kinetic mechanism. This work provides direct evidence that TagE is the wall teichoic acid glycosyltransferase in B. subtilis 168 and provides a strong basis for further studies on the mechanism of wall teichoic acid glycosylation, a largely uncharted aspect of wall teichoic acid biogenesis. | en_US |
dc.language.iso | en | en_US |
dc.subject | glycosylation | en_US |
dc.subject | wall teichoic | en_US |
dc.subject | acid | en_US |
dc.subject | gram-positive | en_US |
dc.subject | bacteria | en_US |
dc.title | Glycosylation of Wall Teichoic Acids in Gram-Positive Bacteria | en_US |
dc.contributor.department | Biochemistry | en_US |
dc.description.degreetype | Thesis | en_US |
dc.description.degree | Doctor of Philosophy (PhD) | en_US |
Appears in Collections: | Digitized Open Access Dissertations and Theses |
Files in This Item:
File | Description | Size | Format | |
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Allison_Sarah_E_2011Apr_Phd.pdf | 2.65 MB | Adobe PDF | View/Open |
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