Anaerobic hydrogen production by mixed cultures is a complex process, and information about its kinetic analysis is sparse. In this study, an integrated approach with the weighted nonlinear least-squares analysis and accelerating genetic algorithm is proposed to evaluate the kinetic parameters of biohydrogen production from a sucrose by mixed anaerobic cultures. The weighted nonlinear least-squares analysis is used to calculate the differences in gaseous and aqueous product concentrations between the predicted and the measured results, while the accelerating genetic algorithm is utilized to optimize the objective function by minimizing the total sum of the squared weighted errors. The kinetic parameters for specific maximum substrate uptake rate, substrate uptake affinity constant, yield coefficient are calculated with this approach, and are validated by the results of the independent experimental results reported in literature. This integrated approach is effective and rapid to estimate the anaerobic hydrogen production kinetics by mixed cultures. Copyright (C) 2013, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.