The activity of the enzyme tryptophanase in the enteric environment was investigated to elucidate the significance of the enzyme in the metabolism of Escherichia coli. The tryptophanase activity, tryptophan content, and indole concentration as well as the numbers of E. coli were determined in the intestinal and fecal contents of conventional, germ-free, and monocontaminated axenic laboratory mice. Increasing the tryptophan content of the diet of mice having a conventional microflora increased the tryptophanase activity of the enteric microflora by a factor of almost 2 but did not increase the numbers of E. coli either absolutely or relative to other facultative enteric coliforms. In the enteric environment, E. coli is responsible for very little tryptophanase activity, a fraction calculated to be less than 0.02%. The values for the experimental parameters were much the same in the contents of the cecum and in the fecal material.