Isolation and characterization of Salmonella typhimurium glyoxylate shunt mutants

Abstract

Growth of Salmonella typhimurium on acetate as a sole carbon source requires expression of the glyoxylate shunt; however, the genes for the glyoxylate shunt enzymes have not been previously identified in S. typhimurium. In this study, we isolated transposon insertions in the genes for the two unique enzymes of this pathway, aceA (isocitrate lyase) and aceB (malate synthase). The aceA and aceB genes were located at 89.5 min on the S. typhimurium genetic map. Genetic linkage to nearby loci indicated that the relative gene order is purDJ metA aceB aceA. Transposon insertions in aceB were polar on aceA, suggesting that the genes form an operon transcribed from aceB to aceA. Transcriptional regulation of the aceBA operon was studied by constructing mini-Mu d(lac Kan) operon fusions. Analysis of these fusions indicated that expression of the aceBA operon is regulated at the level of transcription; the aceBA genes were induced when acetate was present and repressing carbon sources were absent. Although glucose represses expression of the aceBA operon, repression does not seem to be mediated solely by cyclic AMP-cyclic AMP receptor protein complex. Mutants with altered regulation of the aceBA operon were isolated.