Escherichia coli wild-type cells form constitutively the enzyme phospho-beta-glucosidase A, which has a high affinity for phosphorylated aromatic beta-glucosides and a low affinity for phosphorylated beta-methyl-glucoside. Phospho-beta-glucosidase B and beta-glucoside permease I are formed in aromatic beta-glucoside-fermenting mutants. Mutants lacking phospho-beta-glucosidases A and B have been isolated. These mutants showed a reduced rate of inducibility of the beta-glucoside permease I. The restoration of phospho-beta-glucosidase A or B activity resulted in an increased rate of induction of the beta-glucoside permease I. The presence of the phospho-beta-glucosidases was not required for the constitutive biosynthesis of the beta-glucoside permease. Mutants selected for growth on beta-methyl-glucoside as carbon source showed an increased level of constitutive phospho-beta-glucosidase A activity. Gene bglD, the structural gene for phospho-beta-glucosidase A, was mapped between the pyrE locus and the cluster bgl loci, whereas bglE, the regulatory site determining the hyperproduction of phospho-beta-glucosidase A, was mapped between the bgl and ilv clusters. The bglE locus appears to have a regulatory effect on the expression of the bglD gene.