The use of enzymes to catalyze chemical reactions has increased these recent years. Several models have been developed to express the kinetics over these biocatalysts. The most well-known of them, Michaelis-Menten, is used when only one substrate adsorbs on the enzyme. In the case of the esterification reaction, i.e., bimolecular system, a more complex kinetic model such as the Ping-Pong Bi-Bi should be applied. The use of such advanced models is essential for reactor scaleup and to optimize production. However, these models usually do not consider the reaction temperature. To fill this gap, a Ping-Pong Bi-Bi model was developed to produce butyl levulinate from the esterification of levulinic acid over an immobilized enzyme, Novozym®435. Microfluidic technology was used to ensure ideal mixing conditions. The Ping-Pong model, considering inhibition mechanisms, fits the experimental concentrations. ePC-SAFT equation of state was used to estimate the equilibrium constants.