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High-level expression and characterization of recombinant acid urease for enzymatic degradation of urea in rice wine

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Abstract

Ethylcarbamate, a carcinogenic compound, is formed from urea and ethanol in rice wine, and enzymatic elimination of urea is always attractive. In the present work, we amplified the acid urease gene cluster ureABCEFGD from Lactobacillus reuteri CICC6124 and constructed robust Lactococcus lactis cell factories for the production of acid urease. The titer of the recombinant acid urease was increased from 1,550 to 11,560 U/L by optimization of the cultivation process. Meanwhile, the enzyme showed satisfied properties toward urea elimination in the rice wine model system. By incubating the enzyme (50 U/L) at 20 °C for 60 h, about 95.8 % of urea in rice wine was removed. Interestingly, this acid urease also exhibited activity toward ethylcarbamate. The results demonstrated that this recombinant acid urease has great potential in the elimination of urea in rice wine.

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Acknowledgments

We appreciate Professor Byong Lee at Jiangnan University for his discussion and revision. This work was financially supported by the Major State Basic Research Development Program of China (973 Program, 2012CB720802 and 2012CB720806), the National High Technology Research and Development Program of China (863 Program, 2011AA100905), Program for Changjiang Scholars and Innovative Research Team in University (no. IRT1135), the National Science Foundation for Post-doctoral Scientists of China (2013 M540414), the Jiangsu Planned Projects for Postdoctoral Research Funds (1301010B), and the 111 Project.

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Authors and Affiliations

  1. Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, Wuxi, 214122, China

    Yuqing Yang, Zhen Kang, Jianli Zhou, Jian Chen & Guocheng Du

  2. The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China

    Yuqing Yang, Zhen Kang & Jianli Zhou

  3. School of Biotechnology, Jiangnan University, Wuxi, 214122, China

    Yuqing Yang, Zhen Kang, Jianli Zhou, Jian Chen & Guocheng Du

  4. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi, 214122, China

    Jian Chen

  5. The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China

    Guocheng Du

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  1. Yuqing Yang
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  2. Zhen Kang
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  3. Jianli Zhou
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  4. Jian Chen
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Correspondence to Zhen Kang or Guocheng Du.

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Yang, Y., Kang, Z., Zhou, J. et al. High-level expression and characterization of recombinant acid urease for enzymatic degradation of urea in rice wine. Appl Microbiol Biotechnol 99, 301–308 (2015). https://doi.org/10.1007/s00253-014-5916-z

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  • DOI: https://doi.org/10.1007/s00253-014-5916-z

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