During growth of Escherichia coli on acetate, the glyoxylate bypass supplies the precursors needed for biosynthesis. The glyoxylate bypass enzyme isocitrate lyase competes with the citric acid cycle enzyme isocitrate dehydrogenase for the available isocitrate. We have studied the control of metabolic flux at this branchpoint by examining the regulatory properties of the enzymes concerned. Isocitrate dehydrogenase is controlled by reversible phosphorylation catalysed by a bifunctional kinase/phosphatase whose activities are regulated by isocitrate, biosynthetic precursors and adenine nucleotides. The flux through isocitrate lyase is mainly controlled by the intracellular concentration of isocitrate. The phosphorylation system responds to the availability of energy and precursors and maintains isocitrate at a concentration high enough to sustain the flux through the glyoxylate bypass necessary for biosynthesis.