title: In vitro formation of bile acids from di- and trihydroxy-5 beta-cholestanoic acid in human liver peroxisomes.
abstract: The conversion of 3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-[3H]cholestanoic acid into cholic acid and 3 alpha,7 alpha-dihydroxy-5 beta-[3H]cholestanoic acid into chenodeoxycholic acid has been studied in subcellular fractions of human liver. The products were separated from the substrates by high-pressure liquid chromatography and identified by combined gas chromatography-mass spectrometry. The highest rates of conversion were found in the light mitochondrial fraction. This fraction also contained the highest amount of the marker enzymes for peroxisomes. The maximal rates of cholic acid and chenodeoxycholic acid formation were 1.3 and 1.8 nmol/mg protein per h, respectively. The presence of KCN in the incubation medium stimulated the formation of bile acids. Peroxisomes were prepared from the light mitochondrial fraction by sucrose-gradient centrifugation. By use of different marker enzymes, it was confirmed that the major part of the activity for cholic acid formation in the light mitochondrial fraction was located in the peroxisomes. It is concluded that liver peroxisomes are important for the oxidative cleavage of the C27 steroid side chain in bile acid formation in man.
mesh_terms: D000118:Acetylglucosaminidase / Q000378:metabolism; D001647:Bile Acids and Salts / Q000096:biosynthesis; D002374:Catalase / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D002635:Chenodeoxycholic Acid / Q000378:metabolism; D002777:Cholestanols / Q000378:metabolism*; D002793:Cholic Acids / Q000096:biosynthesis / Q000378:metabolism*; D003574:Cytochrome c Group / Q000032:analysis; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D008099:Liver / Q000648:ultrastructure*; D008830:Microbodies / Q000378:metabolism*; D008930:Mitochondria, Liver / Q000201:enzymology; D008995:Monoamine Oxidase / Q000378:metabolism; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism; D011190:Potassium Cyanide / Q000494:pharmacology; D014018:Tissue Distribution
chemical_list: C000986:3,7-dihydroxycholestan-26-oic acid; D001647:Bile Acids and Salts; D002777:Cholestanols; D002793:Cholic Acids; D003574:Cytochrome c Group; D002635:Chenodeoxycholic Acid; C003681:3,7,12-trihydroxycoprostanic acid; D002374:Catalase; D008995:Monoamine Oxidase; D009251:NADPH-Ferrihemoprotein Reductase; D000118:Acetylglucosaminidase; D011190:Potassium Cyanide