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Inhibition of Cholesterol Biosynthesis by Bovine Milk, Cultured Buttermilk, and Orotic Acid

Department of Food Science, University of Wisconsin, Madison, Wisconsin 53706

ABSTRACT

The effects of 50 µl volumes of buffer, pasteurized, homogenized bovine milk, cultured buttermilk, or orotic acid solution (73 mg/liter) on the incorporation of [1-¹⁴C] acetate or [5-³ H] mevalonic acid into cholesterol were determined in 3.5 ml rat liver homogenate preparations. Radioactive cholesterol was isolated as the digitonide after saponification of the homogenate followed by extraction of cholesterol with hexane. The digitonide from the control homogenates exposed to [1-¹⁴C] acetate yielded 3497 ± 545 dpm/mg cholesterol (10 replicates) whereas the digitonides from homogenates containing bovine milk, buttermilk, or orotic acid yielded 56 ± 12 dpm/mg cholesterol (six replicates), 59 ± 13 dpm/mg cholesterol (five replicates), or 151 ± 90 dpm/mg cholesterol (six replicates), respectively. This corresponds to an average inhibition of acetate incorporation into cholesterol of 98.4%, 98.3%, or 95.7%. Homogenates exposed to [5-³ H] mevalonic acid yielded digitonides from the control with 98,228 ± 9663 dpm/mg cholesterol (seven replicates), from bovine milk with 96,690 ± 6784 dpm/mg cholesterol (seven replicates), and from orotic acid with 100,475 ± 7483 dpm/mg cholesterol (seven replicates). Mevalonic acid incorporation was not affected statistically by the presence of bovine milk or orotic acid. This indicates the inhibition of cholesterol biosynthesis prior to the formation of mevalonate. Orotic acid concentrations of the bovine milk and buttermilk used were 69 mg/liter and 53 mg/liter, respectively.