Inducible system for the utilization of beta-glucosides in Escherichia coli. II. Description of mutant types and genetic analysis.

Abstract

Two types of mutants obtained by treating beta-gl(+) cells with nitrosoguanidine are described. One type, beta-gl(+)c, is constitutive for the biosynthesis of the aryl beta-glucoside splitting enzyme(s) and for the beta-glucoside permease; the other (beta-gl(+)sal(-)) has lost the capacity to ferment salicin, but has retained the capacity to ferment arbutin and other aryl beta-glucosides. By two successive mutational steps, beta-gl(+)sal(-)c double mutants can be obtained. Determinations of the enzymatic splitting of salicin and p-nitrophenyl beta-glucoside by beta-gl(+)sal(-) cells and extracts showed that these mutants have lost the capacity to split salicin but do split p-nitrophenyl beta-glucoside; they possess the beta-glucoside permease, and in them salicin is a gratuitous inducer for enzyme and permease biosynthesis. Studies on a beta-gl(+) strain, which splits salicin as well as p-nitrophenyl beta-glucoside, have shown that the splitting of salicin is more temperature-sensitive than that of p-nitrophenyl beta-glucoside and other beta-glucosides. Other properties of the two activities are similar. Interrupted mating experiments and cotransduction with P1kc phage showed that the genetic determinants of the beta-glucoside system map between the pyrE and ile loci. Three distinct mutational sites were found and are presumed to have the following functions: beta-glA, a structural gene for an aryl beta-glucoside splitting enzyme; beta-glB, either the structural gene for the beta-glucoside-permease or a regulatory gene; and beta-glC, a regulatory gene (or site). Escherichia coli wild-type strains are of the genotype A(+) B(-) C(+). The beta-gl(+) mutation determining the ability to ferment beta-glucosides is considered to be a permease or regulatory mutation, and the resulting genotype is A(+) B(+) C(+). The beta-gl(+)sal(-) phenotype results from a mutation in the beta-glA gene (genotype A' B(+) C(+)), and the constitutive phenotype results from a mutation in the beta-glC gene, the genotypes A(+) B(+)C(a) and A' B(+)C(a) corresponding to the phenotypes beta-gl(+)c and beta-gl(+)sal(-)c.