Direct penicillin G acylase immobilization by using the self-prepared immobilized metal affinity membrane

• Published in: Journal of membrane science Vol. 380; no. 1; pp. 34 - 40
• Main Authors: Chen, Chih-I, Ko, Yi-Miao, Shieh, Chwen-Jen, Liu, Yung-Chuan
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15.09.2011; Elsevier
• Subjects: activation energy; artificial membranes; benzylpenicillin; biocatalysts; Catalysis; cellulose; chemical reactions; Chemistry; Colloidal state and disperse state; copper; Enzyme immbolization; Enzyme stability test; epichlorohydrins; Exact sciences and technology; General and physical chemistry; Immobilized metal affinity membrane; industrial applications; Kinetic analysis; Membranes; Penicillin G acylase; temperature; Theory of reactions, general kinetics. Catalysis. Nomenclature, chemical documentation, computer chemistry; Enzyme immbolization; Immobilized metal affinity membrane; Kinetic analysis; Enzyme stability test; Penicillin G acylase; Stability; Enzyme; Industrial application; Cellulose; Immobilization; Activation; Copper ion; Storage; Acids; Energy; Chemical reaction; Penicillin; pH; Affinity; Membrane; Kinetics; Kinetic parameter; Resins; Catalyst activity
• ISSN: 0376-7388; 1873-3123
• DOI: 10.1016/j.memsci.2011.06.027

• An immobilized metal affinity membrane (IMAM) with copper ion chelated was built. • PGA directly immobilize on IMAM. • 99% of the residual activity could be retained in the repeated test. • Little activity loss was observed within 40 days of storage. • The direct PGA immobilization on IMAM facilitates its applications in industry. The use of penicillin G acylase (PGA) requires efficient methods for immobilizing the enzyme to yield a biocatalyst with high activity and stability for industrial application. This study constructs the immobilized metal affinity membrane (IMAM) via a series of chemical reactions to sequentially couple epichlorohydrin (EPI), iminodiacetic acid (IDA), and copper ions on the regenerated cellulose membrane. The feasibility of direct PGA immobilization on IMAM was examined. Various conditions were tested for constructing IMAM for PGA immobilization. Under the optimal condition, a copper ion of 492.8 μmol/g with a PGA activity of 11.76 U/g was obtained for IMAM as compared to those for the commercial immobilized metal affinity resin (299.8 μmol/g and 2.99 U/g). The immobilized PGA membrane was stable in the pH range from 7 to 9 and for temperatures from 40 to 60 °C. In addition, 99% of the residual activity could be retained via a 16-times repeated use. Only little activity loss was observed after a 40 days of storage. This study also determines the kinetic parameters V max, K m, and the energy of activation ( E a) for the immobilized PGA membrane.