To develop a commercial fermentation process producing butanol, a kinetic model describing the metabolism in an acetone-butanol-ethanol (ABE)-producing by Clostridium acetobutylicum ATCC 824, was proposed by modifying the previously proposed metabolic pathways. We used an efficient optimization algorithm combining a genetic algorithm and the Levenberg-Marquardt algorithm in order to estimate the kinetic parameters of the model with experimental data obtained from a batch fermentor because of the complexity of the model equations. To evaluate the kinetic parameters estimated with the fermentation experiments using C. acetobutylicum ATCC824, an additional experiment was carried out using the modified clostridium, where pathway deletion was introduced. Thus, this kinetic study will be contributed to designing C. acetobutylicum for enhanced ABE production.