Immobilization of lipase from Candida rugosa on synthesized hydrogel for hydrolysis reaction

Novel hydrogels based on N-acryloyl-tris(hydroxymethyl) aminomethane (NAT), 2-aminoethyl methacrylate (AEMA) and N,N′-methylenebisacrylamide (BIS) were prepared and applied for immobilization of lipase. Two procedures were previously performed on the hydrogels as follows: (a) modification with epichlorohydrin (ECH) or 1,4-butanediol diglycidyl ether (BDGE) or (b) grafting with AEMA, using ceric ammonium sulfate as an initiator, and subsequent reaction with ECH. In the hydrolysis reactions, high percentages of lipase activity were reached with products whose enzyme immobilization were performed on both BDGE-containing products and grafted matrices, since the enzyme was bound spaced from the matrices and more likely to interact with the substrate. The best derivatives preserved 86.7–89.5% of the activity corresponding to the soluble enzyme.

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► New rod-shaped hydrogels were prepared by co-polymerization of N-acryloyl-tris(hydroxymethyl) aminomethane, 2-aminoethyl methacrylate and N,N′-methylenebisacrylamide.
► Two procedures were performed on the hydrogels: (a) modification with epichlorohydrin (ECH) or 1,4-butanediol diglycidyl ether (BDGE) or (b) grafting with AEMA and subsequent reaction with ECH.
► The oxirane-containing hydrogels reacted with Lipase rugose allowing its immobilization in all cases.
► The enzyme hydrolytic activity of the final products was determined from the hydrolysis of p-nitrophenyl palmitate.
► High percentages of activity were reached in those products in which the enzyme was immobilized on BDGE-containing matrices and grafted matrices.