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Properties of RNase A Immobilized on Magnetic Poly(2‐hydroxyethyl methacrylate) Microspheres
Author(s) -
Horák Daniel,
Rittich Bohuslav,
Šafář Jan,
Španová Alena,
Lenfeld Jiří,
Beneš Milan J.
Publication year - 2001
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp0100171
Subject(s) - cyanuric chloride , rnase p , chemistry , methacrylate , adsorption , nuclear chemistry , copolymer , (hydroxyethyl)methacrylate , chromatography , immobilized enzyme , rna , polymer chemistry , enzyme , organic chemistry , biochemistry , gene , polymer
Magnetic hydrogel microspheres 1.5 μm in size were prepared by dispersion copolymerization of 2‐hydroxyethyl methacrylate and ethylene dimethacrylate in the presence of magnetite, which formed the core of the particles. RNase A was coupled to the particles by the cyanuric chloride method. Gel electrophoresis of plasmid DNA pUC 19 (contaminated by bacterial RNA) confirmed RNA degradation with the immobilized enzyme. The effect of temperature and pH on the relative activity of immobilized RNase A was estimated after incubation of the samples at different temperatures (30−80 °C) and pH (4.0−8.0). Maximum relative activity was observed at 70 °C and pH 6.5. The matrices based on magnetic poly(HEMA) had a low tendency to adsorb RNA.