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 inJournal of microbiology and biotechnology Vol. 19; no. 12; pp. 1650 - 1655
Main Authors Kim, H.S., Seoul National University, Seoul, Republic of Korea, Kim, J.Y., Seoul National University, Seoul, Republic of Korea, Park, M.S., Anyang Technical College, Anyang, Republic of Korea, Zheng, Hua, Seoul National University, Seoul, Republic of Korea, Ji, G.E., Seoul National University, Seoul, Republic of Korea
Format Journal Article
LanguageEnglish
Published Seoul Korean Society for Applied Microbiology 01.12.2009
한국미생물·생명공학회
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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.
Bibliography:A50
2010002488
ObjectType-Article-1
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content type line 23
G704-000169.2009.19.12.002
ISSN:1017-7825
1738-8872
DOI:10.4014/jmb.0904.04053