Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Cloning and Expression of $\beta$-Glucuronidase from Lactobacillus brevis in E. coli and Application in Bioconversion of Baicalin and Wogonoside

  • Kim, Hyun-Sung (Interdisciplinary Program in Genetic Engineering, Seoul National University) ;
  • Kim, Jin-Yong (Department of Food and Nutrition, Research Institute of Human Ecology, Seoul National University) ;
  • Park, Myeong-Soo (Department of Culinary Arts, Anyang Technical College) ;
  • Zheng, Hua (Department of Food and Nutrition, Research Institute of Human Ecology, Seoul National University) ;
  • Ji, Geun-Eog (Department of Food and Nutrition, Research Institute of Human Ecology, Seoul National University)
  • Published : 2009.12.31
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Abstract

The $\beta$-glucuronidase (GUS) gene from Lactobacillus brevis RO1 was cloned and expressed in Escherichia coli GMS407. The GUS gene was composed of 1,812 bp, encoding a 603-amino-acid protein belonging to glycosyl hydrolase family 2 with three conserved domains. The amino acid similarity was higher than 70% with the $\beta$-glucuronidases of various microorganisms, yet less than 58% with the $\beta$-glucuronidase of L. gasseri ADH. Overexpression and purification of the GUS was performed in $\beta$-glucuronidase-deficient E. coli GMS407. The purified GUS protein was 71 kDa and showed 1,284 U/mg of specific activity at optimum conditions of pH 5.0 and $37^{\circ}C$. At $37^{\circ}C$, the GUS remained stable for 80 min at pH values ranging from 5.0 to 8.0. The purified enzyme exhibited a half-life of 1 h at $60^{\circ}C$ and more than 2 h at $50^{\circ}C$. When the purified GUS was applied to transform baicalin and wogonoside into their corresponding aglycones, $150\;{\mu}M$ of baicalin and $125\;{\mu}M$ of wogonoside were completely transformed into baicalein and wogonin, respectively, within 3 h.

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References

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