Purification and spectral characteristics of cytochrome C-551 in a phthalate metabolizing bacterium / by Mark C. Gunderson.

Author/creator Gunderson, Mark Charles author.
Other author Ito, Takeru, degree supervisor.
Other author East Carolina University. Department of Biology.
Format Theses and dissertations
Production1979.
Description60 leaves, 2 unnumbered leaves of plates : illustrations ; 28 cm
Supplemental ContentAccess via ScholarShip
Subjects

Summary Routine Inspection of the harvested bacterial cells from a phthalate metabolizing bacterium showed a reddish tint which was thought to be due to the presence of cytochromes. This report describes purification and characterization of cytochrome c from this organism. Nutrient broth was chosen as the growth medium since It produced the largest quantities of cytochrome c-551 per gram dry weight of bacteria. It was found that nutrition plays a role in the relative concentrations cytochrome c-551 and cytochrome b-558 In the experimental organism. The French Press was found to be very efficient in liberating the cytochrome c-551 from the bacterial cells. The extracted c-551 was further purified with ammonium sulfate fractionation and Ion-exchange techniques with SP-Sephadex. The major purification resulted from Isoelectric focusing which resulted in 250 fold purification over the original cell free extract. Gel electrophoresis confirmed the purity of the final product was very high. The cytochrome c-551 had reduced maxima at 551, 522, and 416 nm. The oxidized peak was at 408 nm. The protein appeared to be a typical cytochrome c similar to many of the prokaryotic c-type cytochromes and mitochondrial cytochrome c. Evidence of electron transport capabilities of the highly purified enzyme indicated that the isolation procedures had not damaged the enzymatic functions of the cytochrome c-551.
Local noteJoyner-"Presented to the faculty of the Department of Biology ... in partial fulfillment of the requirements for the degree Master of Science in Biology.
General notePresented to the Faculty of the Department of Biology
General noteAdvisor: Takeru Ito
Dissertation noteM.S. East Carolina University 1979
Bibliography noteIncludes bibliographical references (leaves 59-60).
Genre/formAcademic theses.
Genre/formAcademic theses.
Genre/formThèses et écrits académiques.

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