The influence of high concentrations of ferric ions on the biochemical activity of Acidithiobacillus ferrooxidans was studied using intact cells. The specific oxidation rate of ferrous ions decreased with increasing ferric ion concentration. Lineweaver-Burk plots revealed typical competitive inhibition kinetics, because the slopes varied with the ferric ion concentration. A linear relationship between the slope and the square of the ferric ion concentration revealed that the iron-oxidizing enzyme system of A. ferrooxidans was competitively inhibited by about two molecules of ferric ion. The kinetic equation based on this inhibition model agreed with the experimental observation at a high ferric ion concentration where the bacterium is usually exposed in bioleaching and biooxidation plants.