Abstract
An extra-mitochondrial system prepared from germinating peanut cotyledons by differential centrifugation and ammonium sulfate precipitation is able to catalyze the conversion of C14-labeled palmitic acid to C14-labeled acetyl-CoA, in the presence of CoA, ATP, DPN and Mg (9). This system is found in the supernatant fraction left after centrifugation of the mitochondria at 10,000 X g for 30 minutes. Of course this fraction is heterogeneous containing some mitochondrial debris, microsomes, as well as soluble proteins. The question of intracellular localization of the enzymes catalyzing this ,6-oxidation activity was raised (9) and 3 possibilities were advanced: A) The /3-oxidation activity of this in vitro extra-mitochondrial fraction is derived from ruptured mitochondria; B) The /8-oxidation activity in the 10,COO X g supernatant is indeed of extra-mitochondrial origin, i.e., microsomal, hyaloplasmic or both. C) In vivo ,6-oxidation activity is present both inside and outside the mitochondria. During the course of this study we were able to show that in vitro, the activation of long chain fatty acids is particulate, while the conversion of the activated acids into acetyl-CoA is catalyzed by very soluble enzymes.
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