The requirements for cysteine of naturally occurring cysteine auxotrophs of Escherichia coli were determined in a defined liquid medium. Maximal growth was obtained in the presence of cysteine concentrations between 20 and 250 mg/l. At concentrations below 20 mg/l growth of the auxotrophs, but not the prototrophic control, was suboptimal in this system. In the presence of cysteine concentrations in excess of 250 mg/l, growth of both auxotrophic and prototrophic E. coli was inhibited with lower growth yields, a decreased specific growth rate and an extended lag phase being observed. These effects were minimised in the presence of 2 mM L-leucine, L-isoleucine and L-valine.