Kresheck, Gordon C.||Albers, Robert J.||Kevill, Dennis N.
M.S. (Master of Science)
Department of Chemistry
Bovine milk alkaline phosphatase has been isolated in a highly purified form by using a 0.1 M potassium acetate gradient at pH 8.5 with diethylaminoethy1-cellulose ion exchange chromatography. The material, obtained in low yields, represented a four fold higher degree of purification than for Morton's most active fraction. In order to obtain purified milk alkaline phosphatase at least two rechromatography fractionations must be performed. In two pilot studies and a preparative study, the enzymatic activity appeared between two non-resolvable protein peaks. Based on the molecular weights of the last preparative rechromatography fractionation, the last pilot rechromatography, and specific peaks of an ion exchange chromatography of a sample obtained from preparative gel electrophesis, the final isolated material from the preparative chromatography fractionation is a composite of the two non-resolvable protein fractions. Recombination and reactivation studies were performed when the double protein pattern was obtained. No signs of any increase in activity resulted, suggesting that the two non-resolvable protein fractions do not represent a sub-unit system, or else the proper conditions for activation or reactivation to occur were not achieved if sub units are involved.
Berg, Thor F., "Chromatographic studies in the purification of bovine milk alkaline phosphatase" (1969). Graduate Research Theses & Dissertations. 1798.
Northern Illinois University
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