Determination of Coenzyme A using Phosphotransacetylase of Streptococcus faeculis

Abstract

It was demonstrated that phosphotransacetylase (acetyl-CoA : orthophosphate acetyltransferase, EC 2.3.1.8.) of Streptococcus faecalis R (ATCC 8043,IFO 3181) could be used for the determination of CoA, thru phosphotransacetylase reaction coupled with the arsenolytic cleavage of acetyl-CoA to acetylarsenate and CoA. The specific activity of the partialy purified enzyme preparation was found to be about three times as high as that of Escherichia coli 26 comparing with each other at the similar purification stages. In the reaction system containing cysteine oxidized CoA or glutathione-(S-S)-CoA was found to serve as substrates of the phosphotransacetylase reaction, whereas precursors of CoA such as pantetheine -4'-hosphate, pantethine-4'-phosphate, 3'-dephospho CoA and pantothenate had shown nc catalytic activity. Both ATP and ADP were found to be inhibitory to the reaction.