PURIFICATION AND CHARACTERIZATION OF FORMYL-COENZYME-A TRANSFERASE FROM OXALOBACTER-FORMIGENES

Formyl-coenzyme A (formyl-CoA) transferase was purified from Oxalobacter formigenes by high-pressure liquid chromatography with hydrophobic interaction chromatography and by DEAE anion-exchange chromatography. The enzyme was a single entity on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel permeation chromatography (M(r), 44,000). It had an isoelectric point of 4.7. The enzyme catalyzed the transfer of CoA from formyl-CoA to either oxalate or succinate. Apparent K(m) and V(max) values, respectively, were 3.0 mM and 29.6 μmol/min per mg for formyl-CoA with an excess of succinate. The maximum specific activity was 2.15 μmol of CoA transferred from formyl-CoA to oxalate per min per mg of protein.