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Partial Purification of the B-Esterase of Bovine Milk

Department of Dairy Science Washington State University Pullman

ABSTRACT

Tenfold purification of the B-esterase (1) of bovine milk has been obtained by adsorption of the enzyme on a Mg(OH)₂ suspension, followed by decomposition of the Mg(OH)₂ to a soluble carbonate with carbon dioxide under pressure and removal of ions by dialysis.

Details of the purification procedure were as follows: Milk known to be high in B-esterase activity was selected. It was withdrawn from the glass jar at the milking parlor into a brown glass bottle, immediately placed in ice water, brought to the laboratory, and gassed with nitrogen (2). It was stored in an atmosphere of nitrogen, at 2–5· C., until the following day. All subsequent operations, except assays for enzyme activity and nitrogen, were conducted in the cold room (2–5· C.). One hundred and sixty milliliters of gravity skim, obtained from the above milk, were added to 40 ml. of Mg(OH)₂ suspension (3) in a 250-ml. centrifuge bottle and shaken vigorously for 30 sec.