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ABSTRACT
Hyperuricaemia is a serious purine metabolism disorder which may lead to gout, nephrolithiasis and other serious health problems. As a key enzyme responsible for the hydrolysis of uric acid to allantoin in the purine degradation pathway, uricase, especially recombinant uricase, shows great potential in the therapy of hyperuricaemia. In this paper, baboon uricase gene attached with Trx and hexahistidine tags was cloned and expressed in Escherichia coli Rosetta (DE3). The target protein formed a soluble structure in the cytoplasm at 25°C, with a gradual enrichment in the induction time course, reaching its highest level at 6 h. After purification and maturation, the final yield of mature baboon uricase was 136.0 mg/L with enzyme activity of 17.93 U/mg. The Km of recombinant uricase was 6.15 7mu;mol/L. And the optimum temperature and pH of the prepared uricase were 37 ⪚C and 8.2, respectively. MALDI-TOF-MS/MS characterization confirmed the prepared protein was identical with the native baboon uricase. These results showed that the baboon uricase gene had been expressed effectively in E. coli and the obtained protein possessed promising enzymatic properties, which gives a new candidate for the therapy of hyperuricaemia.