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Thesis (M.S.)—University of Nebraska—Lincoln, 1968. Department of Biochemistry and Nutrition.


Copyright 1968, the author. Used by permission.


Ovalbumin constitutes about 50% of the protein of egg white. In spite of the fact that it may be crystallized, it is not homogeneous. In spite of the fact that it may be crystallized, it is not homogeneous. It consists of three components, A1, A2, and A3 which comprise 80, 14, and 4 percent of ovalbumin respectively. These may be separated by electrophoresis and column chromatography using CMC as a cation-exchanger.

Due to the small amounts of A2 and A3 which may be obtained by direct chromatography of ovalbumin, a method is needed to provide increased quantities of these fractions in order to enable a detailed study of the properties of these proteins.

A method employing batch fractionation for the removal of A1 thereby increasing the amounts of A2 and A3 which may be place on a column will be investigated. Perlmann has reported the conversions of A­1 to A­2 and A­2 to A­3 by the enzymic removal of one or two phosphorus atoms, respectively. This method will be investigated as a means of preparing larger quantities of A­2 and A­­3. Sulfhydryl and phosphorus determinations together with electrophoretic analysis of the A­2 and A­­3 fractions will be conducted to ascertain whether the proteins prepared by dephosphorylation are identical to those isolated from purified ovalbumin by CMC chromatography.

Preliminary studies of thermal denaturation at various protein concentrations, pH, temperature, and duration of heating will be conducted to determine the heat stability of the individual proteins. Sulfhydryl and electrophoretic analysis of the denatured proteins will be compared to the values and mobilities found for the native state.

Advisor: Robert M. Hill