Dissociation of Dimeric 6-Hydroxymellein Synthase, a Polyketide Biosynthetic Enzyme in Carrot Cell Extracts, with Loss of Keto-Reducing Activity

6-Hydroxymellein synthase, an inducible polyketide biosynthetic enzyme in carrot cell extracts, is composed of two identical subunits, and the homodimer is dissociated to monomeric peptides under high-ionic-strength conditions with loss of the synthase activity. Appreciable radioactivities were associated with the synthase proteins when the monomer enzyme was incubated with the radiolabeled substrates, acetyl-coenzyme A (CoA) and malonyl-CoA. Therefore, it appeared that the synthase does not lose the ability of binding the substrate even after the dissociation to monomers. The monomeric synthase liberated triacetic acid lactone as the derailment product instead of 6-hydroxymellein from the enzyme-attached triketomethylene chain which is the immediate precursor of an NADPH-dependent keto-reducing reaction involved in 6-hydroxymellein biosynthesis. These observations strongly suggest that the monomeric synthase retains the ability of ketomethylene chain elongation by the condensation of acyl-CoAs, but is lacking in an NADPH-dependent keto-reducing activity toward the triketide intermediate. Results obtained in the present experiments imply that the catalytic domain of acyl-CoA condensation is able to associate with that of keto reduction, possibly belonging to another subunit, only in the homodimeric structure to organize the multicatalytic reaction center.