Purification of a diketo acid hydrolase from rat liver and its use for the enzymic determination of 3,5-dioxohexanoate (triacetate).
1. A diketo acid hydrolase was purified about 100-fold from rat liver and some of its properties were studied. 2. Comparison of its activity with 3,5-dioxohexanoate (triacetate) and fumarylacetoacetate at various stages during the purification suggests that it is identical with 4-fumarylacetoacetate fumarylhydrolase. 3. The acetoacetate formed on cleavage of 3,5-dioxohexanoate by the hydrolase can be determined either with 3-hydroxybutyrate dehydrogenase or with 3-oxo acid CoA-transferase and 3-hydroxyacyl-CoA dehydrogenase. These reactions form the basis of enzymic assays for 3,5-dioxohexanoate.
D000090:Acetoacetates / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism; D000429:Alcohol Oxidoreductases; D000818:Animals; D002208:Caproates / Q000032:analysis*; D006867:Hydrolases / Q000302:isolation & purification*; D006884:Hydroxybutyrate Dehydrogenase; D008099:Liver / Q000201:enzymology*; D008722:Methods; D051381:Rats; D014166:Transferases
D000090:Acetoacetates; D002208:Caproates; D000429:Alcohol Oxidoreductases; D006884:Hydroxybutyrate Dehydrogenase; D014166:Transferases; D006867:Hydrolases