Enzyme immobilization techniques on poly(glycidyl methacrylate-co-ethylene dimethacrylate) carrier with penicillin amidase as model.
Two types of bead-form macroporous carriers based on glycidyl methacrylate with ethylene dimethacrylate copolymers were used for the immobilization of penicillin amidase either directly or after chemical modification. Direct binding through oxirane groups, which is equally efficient at pH 4.2 and 7, is relatively slow and brings about an activity loss at low enzyme concentrations. The most efficient immobilization was achieved on glutaraldehyde-activated amino carrier, irrespective of whether the amino groups were formed by ammonia or 1,6-diaminohexane treatment of the original oxirane carrier. Hydrazine treatment gave lower immobilization yields. The same is true of the azide method independent of the length of the spacer. Most enzyme activity was preserved by coupling the carbodiimide-activated enzyme to the carrier with alkyl or arylamino groups at the end of a longer substituent. Immobilization on diazo-modified carrier gave average results. Rapid immobilization by a lysine-modified phosgene-treated carrier resulted in an activity loss. It is suggested that multipoint and very tight attachment of the enzyme molecule to the matrix decreased the activity. The immobilized activity is quite stable in solution and very stable upon lyophilization with sucrose.
D000581:Amidohydrolases / Q000378:metabolism; D000641:Ammonia; D002234:Carbodiimides; D004800:Enzymes, Immobilized / Q000378:metabolism; D005976:Glutaral; D006586:Hexanes; D008768:Methylmethacrylates / Q000031:analogs & derivatives; D008956:Models, Chemical; D010399:Penicillin Amidase / Q000378:metabolism; D013451:Sulfonic Acids; D014050:Toluene
D002234:Carbodiimides; D004800:Enzymes, Immobilized; D006586:Hexanes; D008768:Methylmethacrylates; D013451:Sulfonic Acids; C020347:GMA-EDMA; D014050:Toluene; D000641:Ammonia; D000581:Amidohydrolases; D010399:Penicillin Amidase; D005976:Glutaral