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| Expression, Purification and Activity Identification of Urate Oxidase in Escherichia coli |
| ZHU Lei, WANG Qing-min, WU Guo-dong, XUE Tong-tong, SUN Li-xia, WANG Jing-yi |
| Research and Development Institute of Qilu Pharmaceutical Limited Company, Jinan 250100, China |
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Abstract Urate oxidase (uox, C 1.7.3.3) was responsible for the oxidation of uric acid to allantoin.The urate oxidase gene from Aspergillus flavus was cloned into expression vector of pET43.1a. The sequence was confirmed by DNA sequencing and double endonuclease digestion, and transformed to competent cell named JM109. Recombinant strain was induced to express urate oxidase protein,and the target protein was almost the soluble protein after sonication;the uox pure was purified from supernatant after anion and cation column two-step column purification;the activity of pure uox was determined by spectrophotometry. The results showed that: urate oxidase was high level expressed in E. coli, and the ratio of uox to bacterial proteins could reach 50%; expression product was purified by two steps of chromatography and purification method is simple to obtain high purity protein; the protein had the activity of breaking down uric acid in vitro.
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Received: 09 November 2010
Published: 26 April 2011
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