Purification of Uricase by Biospecific Adsorption-Desorption |
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Authors: | F. Batista-viera R. Axén J. Carlsson |
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Affiliation: | 1. Departamento de Bioqulmíca , Facultad de Ouimica , Montevideo, Uruguay;2. Institute of Biochemistry, University of Uppsala , Box 576, S-75123, Uppsala, Sweden |
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Abstract: | A simple procedure for the purification of uricase from bovine kidney is described. The procedure involves the following steps: 1) processing of kidney mince by borate/butanol, 2) ammonium sulphate precipitation, and 3) biospecific adsorption-desorption. The adsorbents were prepared by chemical attachment of urate or xanthine to agarose gel beads. The desorption was performed by a xanthine solution. The adsorption-desorption procedure resulted in an 11 000–12 000-fold purification. The specific activity of the purified uricase was 19.8 U/mg using either “urate” or “xanthine” adsorbent. The recovery was about 70%.The adsorbents were also used for the purification of commercial uricase preparations from hog liver. In this case the purified uricase also possessed a specific activity of 19.8 U/mg. The products were homogenous as judged by gradipore electrophoresis and gel filtration. |
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Keywords: | α‐Glucosidase Chitosan coated polygalacturonic acid (PGA) beads Immobilization |
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