Cloning and Expression of β-Glucuronidase from Lactobacillus brevis in E. coli and Application in Bioconversion of Baicalin and Wogonoside
The β-glucuronidase (GUS) gene from Laetobacillus 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...
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Published in | Journal of microbiology and biotechnology Vol. 19; no. 12; pp. 1650 - 1655 |
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Main Authors | , , , , |
Format | Journal Article |
Language | English |
Published |
Seoul
Korean Society for Applied Microbiology
01.12.2009
한국미생물·생명공학회 |
Subjects | |
Online Access | Get full text |
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Summary: | The β-glucuronidase (GUS) gene from Laetobacillus 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 β-glucuronidases of various microorganisms, yet less than 58% with the β-glucuronidase of L. gasseri ADH. Overexpression and purification of the GUS was performed in β-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℃. At 37℃, 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℃ and more than 2 h at 50℃. When the purified GUS was applied to transform baicalin and wogonoside into their corresponding aglycones, 150 μM of baicalin and 125 μM of wogonoside were completely transformed into baicalein and wogonin, respectively, within 3 h. |
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Bibliography: | A50 2010002488 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 G704-000169.2009.19.12.002 |
ISSN: | 1017-7825 1738-8872 |
DOI: | 10.4014/jmb.0904.04053 |