Cell-Factories are cells that are used for production. To guide the design of efficient cell-factories accurate computational models are required. These models will help predict the effects of gene knockouts, addition of metabolic pathways and changes in the growth conditions on cellular metabolic pathways and thus on the production efficiency of desired metabolites. To explore the central carbon metabolism of two commonly used cell-factories, Escherichia coli and Saccharomyces cerevisiae, we built kinetic models which consider kinetic mechanisms and enzyme regulations. Then, we used omics reference data (e.g., metabolomics and fluxomics) from a single steady-state time point to generate ensembles of the kinetic models. After generating the models, we calculated the kinetic parameters for the Escherichia coli model and compared the calculated values to literature values. In addition, we performed a metabolic control analysis (MCA) to estimate the level of control each enzyme has on the metabolic fluxes inside the cell. The results demonstrate that computational kinetic models are a powerful tool for investigating the metabolic network inside cells. The kinetic parameters calculated for the Escherichia coli model are relatively close to the literature values, which indicates that the model captures well the kinetics of the enzymes. In addition, the MCA enables the investigation of the complex regulation of the central carbon metabolism and we can identify specific enzymes with high control over the metabolic fluxes. This ability is central for directing the design of cell-factories.