Medicina Interna: Acesso Aberto
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Genetic Engineering of Bacteria that can Produce Urate Oxidase

Xin Cheng, Bo Yang, Dong Liu, L Juan He, Gan Chen, Yong Chen, R Fa Huang and Y Sheng Jiang

The urate oxidase gene was cloned into Lactobacillus bulgaria to produce urate oxidase to decompose uric acid and treat hyperuricemia. Using the Candida utilis urate oxidase gene sequences (uricase, E12709) on GenBank, PCR-amplified urate oxidase gene fragments were inserted into the plasmid pMG36e to construct the recombinant plasmid PMG36e-U, which was then electrotransformed into Lactobacillus bulgaria. We used SDS-PAGE to identify urate oxidase in the cell lysates of the genetically engineered bacteria and to measure urate oxidase activity. The urate oxidase gene was PCR-amplified from the Candida utilis genome. The recombinant plasmid PMG36e-U containing the urate oxidase gene was successfully electrotransformed into Lactobacillus bulgaria. The molecular weight of the urate oxidase subunit synthesized by the genetically engineered bacteria was approximately 34 KD based on SDS-PAGE, and the in vitro enzymatic activity from the bacteria preparation was up to 0.33 u/mL. Conclusion: The urate oxidase gene was cloned into Lactobacillus bulgaria and successfully decomposed uric acid.