alexa Genetic Engineering of Bacteria that can Produce Urate Oxidase
ISSN: 2165-8048

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Research Article

Genetic Engineering of Bacteria that can Produce Urate Oxidase

Xin Cheng1,2, Bo Yang1, Dong Liu1, L Juan He1, Gan Chen3, Yong Chen1, R Fa Huang2 and Y Sheng Jiang1*
1Department of Nephrology, The Second Xiangya Hospital of Central South University, Changsha, China
2Ruikang Hospital of Guangxi Traditional Chinese Medical University, China
3Research Center of Medical Chemistry & Chemical Biology, Chongqing Technology and Business University, China
*Corresponding Author : Y. Sheng Jiang
Department of Nephrology
The Second Xiangya Hospital of Central South University
Changsha, 410011 China
Tel: 86- 731-4582381
E-mail: [email protected]
Received March 02, 2012; Accepted August 28, 2012; Published September 04, 2012
Citation: Cheng X, Yang B, Liu D, He LJ, Chen G (2012) Genetic Engineering of Bacteria that can Produce Urate Oxidase. Intern Med 2:114. doi:10.4172/2165-8048.1000114
Copyright: © 2012 Cheng X, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
 

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.

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