A nonuniform distribution model of immobilized cells was established by the studies on immobilized Gluconobacter oxydans and Bacillus cereus living cells. It was compared with uniform distribution model. Effects of the two typical cell distribution on substrate distribution in immobilized biocatalyst, effective rate factor and selective yield were discussed. No obvious difference was observed between the effects of uniform cell distribution and that of nonuniform cell distribution on kinetics of immobilized G. oxydans and B. cereus living cells, which was then confirmed in experiments. The conclusion is also fit for the studies on kinetics of other immobilized biocatalyst system.