Abstract: A generalized mathematical model was developed to simulate the process of simultaneous biochemical reaction and chromatographic separation.The hydrolysis of sucrose to glucose and fructose was chosen as a model reaction to study the coupling process in a 150cm×1cm glass column.It was found that not only sucrose is completely hydrolyzed,but the glucose-rich and fructose-rich products are also separated.Compared to the concentration overload,the volume overload leads to a higher substrate conversion,while there is little difference for the resolution of products.A good agreement between the theoretical and experimental results is reached at the lower sucrose concentrations.