期刊
BIOORGANIC CHEMISTRY
卷 85, 期 -, 页码 159-167出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bioorg.2018.12.026
关键词
beta-xylosidas; Multi-functional hydrolase; Biotransformation; Thermotoga petrophila; Ginsenoside 20(S)-Rg3 preparation
资金
- National Key R & D Program of China [2016YFD0600805]
- National Natural Science Foundation of China [31570565]
- Qing Lan Project
- Natural Science Foundation of Jiangsu Province of China [BK20131423]
- Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD)
A thermostable beta-xylosidase gene Tpexyl3 from Thermotoga petrophila DSM 13,995 was cloned and overexpressed by Escherichia coli. Recombinant Tpexyl3 was purified, and its molecular weight was approximately 86.7 kDa. Its optimal activity was exhibited at pH 6.0 and 90 degrees C. It had broad specificity to xylopyranosyl, arabinopyranosyl, arabinofuranosyl and glucopyranosyl moieties. The beta-xylosidase activity of the recombinant Tpexyl3 was 6.81 U/mL in the LB medium and 151.4 U/mL in a 7.5 L bio-reactor. It was applied to transform ginsenoside extract into the pharmacologically active minor ginsenoside 20(S)-Rg3, which was combined with thermostable beta-glucosidase Tpebgl3. After transforming under optimal condition, the 20 g/L of ginsenoside extract was transformed into 6.28 g/L of Rg3 within 90 min, with a corresponding molar conversion of 95.0% and Rg3 productivity of 1793.49 mg/L/h, respectively. This study is the highest report of a GH3 family glycosidase with arabinopyranosidase activity and also the first report on the high substrate concentration bioconversion of ginsenoside extract to ginsenoside 20(S)-Rg3 by using two thermostable glycosidases.
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