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Cephalosporin C Acylase (CCA), the key biocatalyst in one-step enzymatic production of 7- amino cephalosporanic acid (7-ACA), was immobilized by amino-activated carrier (LX-1000HA) and epoxy-activated carriers (ES-103B, LX-1000EPC), and the activity was assayed. ES-103B carriers showed an advantage than the others. The amount of free enzyme and salt concentration were tested, and the optimum conditions were 1400U/g and 0.9M. In addition, two different methods (by shaker and packed column) were used in CCA immobilization, and the results demonstrated that the former obtained a higher immobilized CCA activity.