Live Tetrahymena thermophila transforms exogenous cholesterol into 7,22-bis, dehydrocholesterol (DHC) by desaturation at positions C7(8) and C22(23) of the cholesterol moiety. In this first report on expression, isolation, characterization, and reconstitution of Tetrahymena's cholesterol desaturases in cell-free extracts, we describe conditions for increasing the expression of both desaturases based on the addition of specific sterols to the culture medium. Reactions performed in vitro, with isolated microsomes, yield only the mono-unsaturated derivatives, 7-DHC and/or 22-DHC.
View Article and Find Full Text PDFThe exceptional nutritional values of milk have long been recognized. However, important drawbacks of milk and dairy products are their relatively high cholesterol contents and their low proportion of polyunsaturated fatty acids. Hypersensitivity to milk proteins can also draw the public away from fluid milk consumption.
View Article and Find Full Text PDFAppl Microbiol Biotechnol
May 2000
Cell-free preparations of Tetrahymena thermophila catalyze the direct desaturation of cholesterol to delta7-dehydrocholesterol (provitamin D3). The activity was isolated in the microsomal fraction from Tetrahymena homogenates. Delta7-desaturase activity was stimulated fivefold by the addition of 6 mM ATP.
View Article and Find Full Text PDFPurpose: The implementation of an in vivo assay to determine the biological activity of human recombinant erythropoietin (Hu-r EPO) is essential. The purpose of this study was to perform and optimize the conditions of an easy in vivo bioassay suitable for routine testing of quality control of Hu-r EPO preparations.
Material And Methods: Normocythemic 8 weeks female mice treated with different Hu-r EPO doses were employed.