Abstract
A porous resin, acrolein–vinyl acetate–divinyl benzene copolymer, was synthesized by radical suspension polymerization of acrolein with vinyl acetate and divinylbenzene in the presence of a pore-creating agent, toluene. A porous polymer carrier containing both anilino and hydroxy groups was obtained by further reaction of the resin with p-nitroaniline, then reduction of the nitro group to amino group. α-Chymotrypsin was immobilized on the porous polymer carrier by the glutaraldehyde–crosslinking method or diazo-coupling method. The activity recovery and relative activity of the immobilized α-chymotrypsin were determined when l-tyrosine ethyl ester and casein were selected as low and high molecular weight substrates, respectively. At the same time, the stability, resistance against inhibition, the Michaelis constant Km, and reusability of the immobilized enzyme were also investigated. The results indicated that the immobilized α-chymotrypsin had not only higher relative activity, but also remarkable stability and better reusability. In addition, the immobilized α-chymotrypsin had stronger resistance against Cu2+ inhibition.
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