The objective of this study was to estimate the first-order intrinsic kinetic constant (k₁) and the liquid-phase mass transfer coefficient (k_c) in a bench-scale anaerobic sequencing batch biofilm reactor (ASBBR) fed with glucose. A dynamic heterogeneous mathematical model, considering two phases (liquid and solid), was developed through mass balances in the liquid and solid phases. The model was adjusted to experimental data obtained from the ASBBR applied for the treatment of glucose-based synthetic wastewater with approximately 500 mg L^–1 of glucose, operating in 8 h batch cycles, at 30 °C and 300 rpm. The values of the parameters obtained were 0.8911 min^–1 for k₁ and 0.7644 cm min^–1 for k_c. The model was validated utilizing the estimated parameters with data obtained from the ASBBR operating in 3 h batch cycles, with a good representation of the experimental behavior. The solid-phase mass transfer flux was found to be the limiting step of the overall glucose conversion rate.