Abstract
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.
Highlights
Hyperuricemia is a direct cause of gout and related diseases and an independent risk factor [1,2,3] for certain kidney and cardiovascular diseases
Urate oxidase synthesis is regulated by the uricase gene and is the critical enzyme for uric acid metabolism
A study of the urate oxidase cDNA homology sequences in humans, apes and other primates found that uricase is composed of four exons
Summary
Hyperuricemia is a direct cause of gout and related diseases and an independent risk factor [1,2,3] for certain kidney and cardiovascular diseases. It is essential to reduce the level of uric acid in blood and tissues to prevent and treat many uric acidrelated diseases. Because the human body cannot synthesize urate oxidase by itself, reducing the level of uric acid requires long-term or even lifelong treatment. Currently various uric acid-lowering drugs cause different degrees of damage to the human body and are not suitable for long-term treatment. A probiotic that can yield a large amount of urate oxidase and be transplanted into the intestines to constantly produce urate oxidase to catalyze uric acid would have important economic and ecological value. We cloned the urate oxidase gene into Lactobacillus bulgaria to assess urate oxidase production and activity as an alternative drug therapy for hyperuricemia
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