Purification and Characterization of Bovine Liver Uridine Phosphorylase

dc.contributor.advisor Kline, Paul en_US
dc.contributor.author Avant, Jamie Danielle en_US
dc.contributor.committeemember Dunlap, Norma en_US
dc.contributor.committeemember Burden, Donald en_US
dc.contributor.department Chemistry en_US
dc.date.accessioned 2014-08-28T18:34:09Z
dc.date.available 2014-08-28T18:34:09Z
dc.date.issued 2014-06-21 en_US
dc.description.abstract Uridine phosphorylase (UPP) is an enzyme that catalyzes the reversible phosphorylytic cleavage of uridine to uracil and αribose–1–phosphate.1 A strong interest has been expressed in the enzyme in recent years because of the recognition of its many medicinal applications. Naturally occurring uridine has been shown to exhibit a cytoprotective effect against the toxicity associated with chemotherapy treatments for both oncological and infectious diseases.2 Inhibition of UPP drastically slows the breakdown of uridine and therefore raises plasma uridine levels in mammals. Determining a purification scheme is the first step to acquiring a pure enzyme that can be studied in the pursuit of the development of stronger and more specific UPP inhibitors. The purification scheme outlined in this thesis consisted of the following steps: ammonium sulfate precipitation, Mono Q ion exchange chromatography, Sephacryl S100 size exclusion chromatography, and hydroxyapatite chromatography. UPP was purified 64–fold with 4.01% final yield. The initial extract showed a specific activity of 0.41 nmoles/min/mg, which increased to 26.51 nmoles/min/mg after the final purification step. Additionally, the purified enzyme was found to exhibit cleaving activity for uridine, thymidine and inosine in the ratio of 5:1:0. en_US
dc.description.degree M.S. en_US
dc.identifier.uri http://jewlscholar.mtsu.edu/handle/mtsu/4272
dc.publisher Middle Tennessee State University en_US
dc.subject Beef en_US
dc.subject Bovine en_US
dc.subject Liver en_US
dc.subject Nucleoside en_US
dc.subject Phosphorylase en_US
dc.subject Uridine en_US
dc.subject.umi Biochemistry en_US
dc.thesis.degreegrantor Middle Tennessee State University en_US
dc.thesis.degreelevel Masters en_US
dc.title Purification and Characterization of Bovine Liver Uridine Phosphorylase en_US
dc.type Thesis en_US
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