The Formation of Bile Acids from Cholesterol. THE CONVERSION OF 5&szligbeta;-CHOLESTANE-3agr,7agr,12agr-TRIOL-26-OIC ACID TO CHOLIC ACID VIA 5&szligbeta;-CHOLESTANE-3agr,7agr,12agr,24xgr-TETRAOL-26-OIC ACID I BY RAT LIVER

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The Formation of Bile Acids from Cholesterol

THE CONVERSION OF 5 ${beta}$ -CHOLESTANE-3 $agr$ ,7 $agr$ ,12 $agr$ -TRIOL-26-OIC ACID TO CHOLIC ACID VIA 5 ${beta}$ -CHOLESTANE-3 $agr$ ,7 $agr$ ,12 $agr$ ,24 $xgr$ -TETRAOL-26-OIC ACID I BY RAT LIVER

Takako Masui ¹ and Ezra Staple ¹

From the ¹ From the Department of Biochemistry, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104

The synthesis of 5ß-cholestane-3 $agr$ ,7 $agr$ ,12 $agr$ ,24 $xgr$ -tetraol-26-oicacid-¹⁴C and the separation of 5ß-cholestane-3 $agr$ ,7 $agr$ ,-12 $agr$ ,24 $xgr$ -tetraol-26-oicacids I and II are described.

The conversion of 5ß-cholestane-3 $agr$ ,7 $agr$ ,12 $agr$ -triol-26-oicacidto 5ß-cholestane-3 $agr$ ,7 $agr$ ,12 $agr$ ,24 $xgr$ -tetraol-26-oic acid I was catalyzedby rat liver mitochondria supplemented with 105,000 x g supernatantfraction, adenosine triphosphate, MgCl₂, flavin adenine dinucleotide,and coenzyme A. Cell fractionation and requirement of cofactorsfor the same reaction were examined.

5ß-Cholestane-3 $agr$ ,7 $agr$ ,12 $agr$ ,24 $xgr$ -tetraol-26-oicacid I which was derived from the incubation of 5ß-cholestane-3 $agr$ ,7 $agr$ ,12 $agr$ -triol-26-oicacid was further oxidized to cholic acid with 105,000 x g supernatantfraction of rat liver, ATP, MgCl₂, glutathione, nicotinamideadenine dinucleotide, and CoA.

The conversion of synthetic 5ß-cholestane-3 $agr$ ,7 $agr$ ,12 $agr$ ,24 $xgr$ -tetraol-26-oicacid-¹⁴C was examined with the use of different cell fractionsof rat liver. The formation of cholic acid from 5ß-cholestane-3 $agr$ ,7 $agr$ ,12 $agr$ ,24 $xgr$ -tetraol-26-oicacid was catalyzed by either mitochondria or 105,000 x g supernatantwith NAD⁺ or NADP⁺.

Submitted on January 18, 1966

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