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KMID : 0545120090190121650
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Journal of Microbiology and Biotechnology
2009 Volume.19 No. 12 p.1650 ~ p.1655
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Cloning and Expression of ?beta-Glucuronidase from Lactobacillus brevis in E. coli and Application in the Bioconversion of Baicalin and Wogonoside
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Kim Hyun-Sung
Kim Jin-Yong
Zheng Hua
Ji Geun-Eog
Park Myeong-Soo
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Abstract
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Beta-glucuronidase (GUS) gene from Lactobacillus brevis RO1 was cloned and expressed in Escherichia coli GMS407. The GUS gene was composed of 1812 bp encoding a 603-amino-acid protein belonging to the glycosyl hydrolase family 2 with three conserved domains. It showed more than 70% of amino acid similarity with the ?beta--glucuronidases of various microorganisms, but less than 58% with that of L. gasseri ADH. Overexpression and purification of GUS was performed in ?beta-glucuronidase-deficient E. coli GMS407. The purified GUS protein was 71 kDa and showed 1284 U/mg of specific activity at optimum condition with pH 5.0 and 37°C. At 37°C, GUS was 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°C and greater than 2 h at 50°C. When purified GUS was applied to transform baicalin and wogonoside, 150 μM of baicalin and 125 μM of wogonoside were completely transformed into the corresponding aglycones, baicalein and wogonin, respectively, within 3 h.
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KEYWORD
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baicalin, baicalein, beta-Glucuronidase, Lactobacillus, wogonin, wogonoside
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