Purification of five components from Clostridium thermoaceticum which catalyze synthesis of acetate from pyruvate and methyltetrahydrofolate. Properties of phosphotransacetylase.
A five-component enzyme system which catalyzes synthesis of acetylphosphate from methyltetrahydrofolate (CH3THF) plus pyruvate has been purified from the homoacetate-fermenting bacterium, Clostridium thermoaceticum. One of the components was identified as the low potential electron carrier, ferredoxin, and the other 4 protein components have been designated F1, F2, F3, and F4. F1, F2, and F4 have been purified to homogeneity and, as estimated by gel filtration, have native molecular weights of 88,100, 58,900, and 255,000, respectively, while the subunit molecular weights obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis are 20,000, 25,500, and 120,000, respectively. F3 contains 3 to 4 protein bands and has not been characterized with respect to molecular weights. Acetylphosphate synthesis by the purified system is optimal at pH 6.0 and 65 degrees C and requires ATP, CoA, and, to a lesser extent, thiamin pyrophosphate and Fe2+. S-Adenosylmethionine is not required. The F1 component has been identified as phosphotransacetylase and in its absence, the product is acetyl-CoA. Some properties of the phosphotransacetylase are presented. A scheme is given indicating present views of the functions of the individual components.
D000085:Acetates / Q000378:metabolism*; D000123:Acetyltransferases / Q000378:metabolism*; D003013:Clostridium / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000302:isolation & purification / Q000378:metabolism*; D010709:Phosphate Acetyltransferase / Q000302:isolation & purification / Q000378:metabolism*; D011773:Pyruvates / Q000378:metabolism*; D019289:Pyruvic Acid; D013763:Tetrahydrofolates / Q000378:metabolism*
D000085:Acetates; D009097:Multienzyme Complexes; D011773:Pyruvates; D013763:Tetrahydrofolates; D008345:Manganese; D019289:Pyruvic Acid; D000123:Acetyltransferases; D010709:Phosphate Acetyltransferase; D008274:Magnesium; C005984:5-methyltetrahydrofolate