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Total Synthesis of Acetate from CO₂: Methyltetrahydrofolate, an Intermediate, and a Procedure for Separation of the Folates

^a Department of Biochemistry, Case Western Reserve University, Cleveland, Ohio 44106

ABSTRACT

Whole-cell preparations of Clostridium thermoaceticum were exposed to a short pulse of ¹⁴CO₂ under conditions in which double-labeled acetate was synthesized. Radioactive methyltetrahydrofolate monoglutamate, diglutamate, and triglutamates were isolated from extracts of the cells. The radioactivity was found to be exclusively in the five methyl position. The specific activities of the methyltetrahydrofolate derivatives were very high and were in accord with the proposal that methyltetrahydrofolates are the precursors of the methyl of acetate. A new method of separation of folates employing QAE-Sephadex chromatography and a linear gradient with triethylammonium bicarbonate is presented which completely resolves the common folate monoglutamates and, upon freeze-drying, yields salt-free preparations.

¹ Present address: Department of Pathology, Washington University, St. Louis, Mo. 63110.