Abstract
The immobilization ofEscherichia coli penicillin acylase (EC 3.5.1.11) was investigated by radiation-induced polymerization of 2-hydroxyethyl methacrylate at low temperature. A leak-proof composite that does not swell in water was obtained by adding the crosslinking agent trimethylolpropane trimethacrylate to the monomer-aqueous enzyme mixture. Penicillin acylase, which was immobilized with greater than 70% yield, possessed a higherK m value toward the substrate 6-nitro-3-phenylacetamidobenzoic acid than the free enzyme form (K m = 1.7 × 10−5 and 1 × 10−5M, respectively). The structural stability of immobilized penicillin acylase, as assessed by heat, guanidinium chloride, and pH denaturation profiles, was very similar to that of the free-enzyme form, thus suggesting that penicillin acylase was entrapped in its native state into aqueous free spaces of the polymer matrix.
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Boccù, E., Carenza, M., Lora, S. et al. Radiation-induced polymerization for the immobilization of penicillin acylase. Appl Biochem Biotechnol 15, 1–10 (1987). https://doi.org/10.1007/BF02798502
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DOI: https://doi.org/10.1007/BF02798502