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Stabilization of penicillin V acylase from Streptomyces lavendulae by covalent immobilization
Author(s) -
TorresBacete Jesús,
Arroyo Miguel,
TorresGuzmán Raquel,
Mata Isabel de la,
Castillón M Pilar,
Acebal Carmen
Publication year - 2001
Publication title -
journal of chemical technology and biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.64
H-Index - 117
eISSN - 1097-4660
pISSN - 0268-2575
DOI - 10.1002/jctb.403
Subject(s) - chemistry , immobilized enzyme , enzyme , covalent bond , kinetics , biocatalysis , chromatography , penicillin , enzyme kinetics , streptomyces , nuclear chemistry , biochemistry , organic chemistry , catalysis , active site , reaction mechanism , antibiotics , bacteria , physics , quantum mechanics , biology , genetics
Abstract Penicillin V acylase from the actinomycete Streptomyces lavendulae ATCC 13664 has been immobilized to epoxy‐activated acrylic beads (Eupergit C®) by covalent binding. Further linkage of bovine serum albumin after enzyme immobilization was carried out in order to remove the remaining oxirane groups of the support. The obtained immobilized biocatalyst displayed double exponential deactivation kinetics at temperatures below 55 °C, while the native enzyme followed single exponential decay at the same temperatures. We concluded that soluble penicillin acylase was deactivated in one step, whereas the immobilized enzyme showed an enzymatic intermediate state which is highly thermostable. As a consequence of the immobilization process, the enzyme displayed a 10‐fold increase in its half‐life at 40 °C. At this temperature, the enzymatic intermediate state was progressively destabilized as the pH of the medium was increased. Thus, the optimum pH range for the immobilized enzyme preparation was established as being from 7.0 to 8.0. Higher pH values led to quicker enzyme deactivation. © 2001 Society of Chemical Industry