Penicillin is the most widely used antibiotics, due to their great spectrum of antibacterial activity, low toxicity, and in- hibitory effect on bacterial cell wall synthesis and it shares about 19 % of the estimated worldwide antibiotic market (Parmar et al. 2000). Therefore, penicillin G acylase (PGA) that catalyzes the hydrolysis of penicillin G to produce 6- aminopenicillanic acid (6-APA) is very essential for the pro- duction of semisynthetic penicillin. It is also important to cre- ate efficient methods for immobilization of PGA for high ac- tivity and stability to make it viable for industrial applications. Immobilization of PGA on immobilized metal affinity mem- brane (IMAM) showed more PGA activity compared to those immobilized metal affinity resin. Immobilization gave stabil- ity to PGA in the pH range from 7 to 9 and for temperatures from 40 to 60 °C. Immobilization also allowed the enzyme to be reused 16 times and caused very little activity loss after 40 days of storage (Chen et al. 2011).