Immobilization of penicillin G acylase on methacrylate polymers.
Macroporous weak cation-exchange methacrylate polymers were synthesized for the immobilization of penicillin G (Pen G) acylase. The role of certain factors such as pore-generating solvent, cross-linking agent, cross-linking density, and comonomer, in enzyme adsorption and expression was studied. Kerosene was a superior pore-generating solvent to paraffin oil. Ethylene glycol dimethacrylate and acrylic acid served as the best cross-linking agent and comonomer, respectively, in the systems studied. 80.3% of the activity of the enzyme adsorbed onto polymer beads prepared with 0.05 mol of acrylic acid (polymer PM-39) was expressed. Properties of the Pen G acylase, immobilized on PM-39 by adsorption and cross-linking with glutaraldehyde (IME-PM-39) were studied. The optimum pH, optimum temperature and Km of Pen G acylase shifted from 8.0 to 7.5-7.8, 50 degrees C to 55 degrees C and 0.038 mol dm-3 to 2.4-3.0 mol dm-3, respectively, as a result of immobilization on PM-39. IME-PM-39 was used repeatedly for 15 cycles in the production of 6-amino penicillanic acid (6-APA).
Polymer Science and Engineering, Chemical Engineering Division, National Chemical Laboratory, Pune, India.
L
L
Koilpillai
R A
RA
Gadre
S
S
Bhatnagar
R C
RC
Raman
S
S
Ponrathnam
K K
KK
Kumar
G R
GR
Ambekar
J G
JG
Shewale
D002411:Cation Exchange Resins / Q000138:chemical synthesis; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004800:Enzymes, Immobilized / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007648:Kerosene; D008567:Membranes, Artificial; D008689:Methacrylates / Q000138:chemical synthesis / Q000378:metabolism*; D010232:Paraffin; D010397:Penicillanic Acid / Q000138:chemical synthesis / Q000031:analogs & derivatives; D010399:Penicillin Amidase / Q000378:metabolism*; D010400:Penicillin G / Q000378:metabolism; D011108:Polymers
D002411:Cation Exchange Resins; D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D007648:Kerosene; D008567:Membranes, Artificial; D008689:Methacrylates; D011108:Polymers; D010232:Paraffin; D010397:Penicillanic Acid; D010399:Penicillin Amidase; D010400:Penicillin G; C004408:aminopenicillanic acid