Aminoacylase was immobilized on the mycelium cells of Aspergillus oryzae by using ethylenediamine, gelatin and glutylalhyde. The proper immobilized condition was studied by orthogonal experimental design. The immobilized cells with excellent activity and stability for optically resoluting N acetyl DL alanine were obtained. The effects of pH, temperature, ion concentration and substrate concentration on the reactive activity of immobilized cells were studied. The continuous optical resolution of N acetyl DL alanine was investigated respectively in an immobilized cells column (ICC) and in a novel couple immobilized cells bed & membrane reactor(CICBMR). The results indicate that the immobilized cells are suitable for industrial applications.