Kinetic Studies of Dehaloperoxidase-Hemoglobin From Amphitrite Ornata

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dc.contributor.advisor Dr. Clay Clark, Committee Member en_US
dc.contributor.advisor Dr. Edmond Bowden, Committee Member en_US
dc.contributor.advisor Dr. Stefan Franzen, Committee Chair en_US Gilvey, Lauren Brenna Galvagni en_US 2010-04-02T17:53:13Z 2010-04-02T17:53:13Z 2006-09-19 en_US
dc.identifier.other etd-08302006-104458 en_US
dc.description.abstract The focus of this research was to determine how three different factors affect the kinetics of the reaction between Dehaloperoxidase — Hemoglobin (DHP-Hb), a halogenated phenol substrate, and a peroxide co-substrate. The factors examined were the order of addition of the substrate and co-substrate, the pH of the solutions, and the difference between using hydrogen peroxide (H2O2) and m-Chloroperbenzoic acid (MCPBA) as the co-substrates. The order of addition studies were carried out on a stop-flow apparatus. The results presented here demonstrate that the substrate must bind to DHP-Hb prior to the addition of the peroxide co-substrate. The effect of pH was examined using stop-flow and photodiode array spectrometry. These studies show how the pKa of the substrate affects the mechanism of the reaction. The kinetic results actually depend on the pKa of the substrate used. Stop-flow studies and UV-Vis assays were used to compare H2O2 and MCPBA as co-substrates. These studies demonstrate that the use of MCPBA as the co-substrate yields a faster rate of product formation. The use of H2O2 as the co-substrate results in a larger amount of product formation at all pHs studied, with the exception of pH 6.0. en_US
dc.rights I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to NC State University or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. en_US
dc.subject Assay en_US
dc.subject UV-Vis en_US
dc.subject Hemoglobin en_US
dc.subject Dehaloperoxidase en_US
dc.subject Peroxidase en_US
dc.subject Stop-Flow en_US
dc.title Kinetic Studies of Dehaloperoxidase-Hemoglobin From Amphitrite Ornata en_US MS en_US thesis en_US Chemistry en_US

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