Cloning, overexpression and characterization of a thermostable β-xylosidase from Thermotoga petrophila and cooperated transformation of ginsenoside extract to ginsenoside 20(S)-Rg3 with a β-glucosidase
| Autor: | Zhenzhong Wang, Shanshan Zhang, Wei Xiao, Erzheng Su, Jianjun Pei, Linguo Zhao, Jingcong Xie |
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| Rok vydání: | 2019 |
| Předmět: |
Ginsenosides
Bioconversion medicine.disease_cause 01 natural sciences Biochemistry law.invention Thermotoga chemistry.chemical_compound Bacterial Proteins law Drug Discovery Escherichia coli medicine Glycoside hydrolase Cloning Molecular Molecular Biology Biotransformation Thermotoga petrophila Cloning Bacteria 010405 organic chemistry Chemistry beta-Glucosidase Organic Chemistry Recombinant Proteins 0104 chemical sciences 010404 medicinal & biomolecular chemistry Transformation (genetics) Xylosidases Ginsenoside Recombinant DNA |
| Zdroj: | Bioorganic Chemistry. 85:159-167 |
| ISSN: | 0045-2068 |
| Popis: | A thermostable β-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 °C. It had broad specificity to xylopyranosyl, arabinopyranosyl, arabinofuranosyl and glucopyranosyl moieties. The β-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 β-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. |
| Databáze: | OpenAIRE |
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