In Saccharomyces cerevisiae, the expression of invertase, which is the hydrolyzing enzyme of sucrose, is controlled by the presence of monosaccharides, such as glucose and fructose, and referred to as carbon catabolite repression. To date, efforts have been made to identify the mechanism by which cells sense extracellular monosaccharide concentrations and trigger the genes involved in the repression pathway. The aim of the present work was to quantitatively investigate the cellular regulation of invertase expression in the wild-type strain S. cerevisiae CEN.PK113-7D during batch growth containing mixed sugar substrates under different initial conditions. Because of the high frequency and accurate online analysis of multiple components, a tight control of invertase expression could be observed, and threshold concentrations of the monosaccharicles for derepression could be determined to 0.5 gl(-1) for glucose and 2 gl(-1) for fructose. Also, the existence of a hitherto undescribed regulatory state, in which cells regulate invertase expression very precisely and operate over long periods at monosaccharide concentrations lower than the above thresholds, could be demonstrated. All experimental observations could be summarized in a formulation of the cellular regulation scheme of invertase expression. A simple kinetic model could show that the regulation scheme explains the observed behavior very well. Additionally, the model was able to explain consequences of the regulation on the global metabolism.