The Enzymatic Carboxylation of Propionyl Coenzyme A: STUDIES INVOLVING DEUTERATED AND TRITIATED SUBSTRATES

Abstract

Abstract (R)- and (S)-2-deuteriopropionyl coenzyme A of approximately 75% optical purity were prepared by a series of reactions starting from optically pure alanine. With these compounds as substrates for propionyl-CoA carboxylase, the deuterium isotope effect was found to be small and probably secondary. No tritium isotope effect was observed when enzymatically prepared 2-3H-propionyl-CoA was used as substrate. The rate of release of tritium was the same as that of the over-all reaction. Tritium from 3H2O was not incorporated into propionyl-CoA in the absence of the reverse reaction. The data were consistent with a concerted mechanism in which an α-proton was removed simultaneously with carboxylation of propionyl-CoA. By various techniques the reaction was shown to proceed with retention of configuration about the α-carbon.