Phenoxiodinin-5-ium chloride (PC) is a synthetic organic compound possessing a little known type of iodine-containing structure and has been found to possess antibacterial activity. A study was conducted to gain some insight into the mechanism by which it exerted its effect on bacteria. The compound inhibited oxygen uptake and 14CO2 evolution from uniformly labelled glucose-14C by Staphylococcus aureus, under both resting and growing cell conditions. The assimilation of glucose-14C and L-methionine-methyl-14C into cellular material was also affected. A release of 14C-labelled cellular material from whole cells and from spheroplasts stabilized in 0.5 M sucrose occurred in the presence of PC. PC also inhibited succinate and lactate dehydrogenase systems in whole cells and cell membranes of Bacillus megaterium. When sodium malonate, a highly specific inhibitor of succinate dehydrogenase, was substituted for PC, an increased release of 14C-intra-cellular material also occurred. PC possesses the ability to inhibit enzymes located on the cell membrane of S. aureus and B. megaterium.