A Novel Ginsenoside-Transforming α-L-Rhamnosidase from <i>Bifidobacterium</i>: Screening, Characterization and Application
Despite the rapid advancement of glycosidase biotechnology, ginsenoside-transforming rhamnosidases remain underexplored due to a lack of research. In this study, we aimed to bridge this gap by evaluating eight putative rhamnosidases for their ability to transform ginsenosides. Among them, a novel rh...
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2024-12-01
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| author | Chang-Hao Cui Doohang Shin Byung-Serk Hurh Wan-Taek Im |
| author_facet | Chang-Hao Cui Doohang Shin Byung-Serk Hurh Wan-Taek Im |
| author_sort | Chang-Hao Cui |
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| description | Despite the rapid advancement of glycosidase biotechnology, ginsenoside-transforming rhamnosidases remain underexplored due to a lack of research. In this study, we aimed to bridge this gap by evaluating eight putative rhamnosidases for their ability to transform ginsenosides. Among them, a novel rhamnosidase (C118) from <i>Bifidobacterium</i> was identified as being efficient at hydrolyzing ginsenoside Re. This enzyme was expressed well in <i>Escherichia coli</i> and exhibited optimal activity at pH of 6.0 and 45 °C. Protein structural predictions revealed that the potential active hydrophobic area near an active pocket may influence the ginsenoside-transforming activities compared to non-active screened rhamnosidases. This enzyme’s thermal stability exceeded that of the only previously known ginsenoside-transforming rhamnosidase, BD890. Additionally, the <i>k<sub>cat</sub></i>/<i>K<sub>m</sub></i> value of C118 was 1.45 times higher than that of BD890. Using recombinant C118 from <i>E. coli</i>, all ginsenoside Re in a PPT-type ginsenoside mixture (2.25 mg/mL) was converted after 12 h of reaction. To the best of our knowledge, this is the most efficient ginsenoside Re-transforming α-L-rhamnosidase reported to date, enhancing our understanding of rhamnosidase–substrate interactions and potentially improving the efficiency and specificity of the conversion process. These findings offer promising implications for the production of pharmacologically active ginsenosides in the pharmaceutical, cosmetic, and functional food industries. |
| format | Article |
| id | doaj-art-231cca4c3f004da5b81ea0d84b79a2cd |
| institution | Kabale University |
| issn | 2218-273X |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
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| series | Biomolecules |
| spelling | doaj-art-231cca4c3f004da5b81ea0d84b79a2cd2024-12-27T14:13:56ZengMDPI AGBiomolecules2218-273X2024-12-011412161110.3390/biom14121611A Novel Ginsenoside-Transforming α-L-Rhamnosidase from <i>Bifidobacterium</i>: Screening, Characterization and ApplicationChang-Hao Cui0Doohang Shin1Byung-Serk Hurh2Wan-Taek Im3Sempio Fermentation Research Center, Sempio Foods Company, Osong 28156, Republic of KoreaSempio Fermentation Research Center, Sempio Foods Company, Osong 28156, Republic of KoreaSempio Fermentation Research Center, Sempio Foods Company, Osong 28156, Republic of KoreaDepartment of Biotechnology, Hankyong National University, Anseong 17579, Republic of KoreaDespite the rapid advancement of glycosidase biotechnology, ginsenoside-transforming rhamnosidases remain underexplored due to a lack of research. In this study, we aimed to bridge this gap by evaluating eight putative rhamnosidases for their ability to transform ginsenosides. Among them, a novel rhamnosidase (C118) from <i>Bifidobacterium</i> was identified as being efficient at hydrolyzing ginsenoside Re. This enzyme was expressed well in <i>Escherichia coli</i> and exhibited optimal activity at pH of 6.0 and 45 °C. Protein structural predictions revealed that the potential active hydrophobic area near an active pocket may influence the ginsenoside-transforming activities compared to non-active screened rhamnosidases. This enzyme’s thermal stability exceeded that of the only previously known ginsenoside-transforming rhamnosidase, BD890. Additionally, the <i>k<sub>cat</sub></i>/<i>K<sub>m</sub></i> value of C118 was 1.45 times higher than that of BD890. Using recombinant C118 from <i>E. coli</i>, all ginsenoside Re in a PPT-type ginsenoside mixture (2.25 mg/mL) was converted after 12 h of reaction. To the best of our knowledge, this is the most efficient ginsenoside Re-transforming α-L-rhamnosidase reported to date, enhancing our understanding of rhamnosidase–substrate interactions and potentially improving the efficiency and specificity of the conversion process. These findings offer promising implications for the production of pharmacologically active ginsenosides in the pharmaceutical, cosmetic, and functional food industries.https://www.mdpi.com/2218-273X/14/12/1611α-L-rhamnosidaseginsenosidebiotransformationginsenoside Rg1ginsenoside Re |
| spellingShingle | Chang-Hao Cui Doohang Shin Byung-Serk Hurh Wan-Taek Im A Novel Ginsenoside-Transforming α-L-Rhamnosidase from <i>Bifidobacterium</i>: Screening, Characterization and Application Biomolecules α-L-rhamnosidase ginsenoside biotransformation ginsenoside Rg1 ginsenoside Re |
| title | A Novel Ginsenoside-Transforming α-L-Rhamnosidase from <i>Bifidobacterium</i>: Screening, Characterization and Application |
| title_full | A Novel Ginsenoside-Transforming α-L-Rhamnosidase from <i>Bifidobacterium</i>: Screening, Characterization and Application |
| title_fullStr | A Novel Ginsenoside-Transforming α-L-Rhamnosidase from <i>Bifidobacterium</i>: Screening, Characterization and Application |
| title_full_unstemmed | A Novel Ginsenoside-Transforming α-L-Rhamnosidase from <i>Bifidobacterium</i>: Screening, Characterization and Application |
| title_short | A Novel Ginsenoside-Transforming α-L-Rhamnosidase from <i>Bifidobacterium</i>: Screening, Characterization and Application |
| title_sort | novel ginsenoside transforming α l rhamnosidase from i bifidobacterium i screening characterization and application |
| topic | α-L-rhamnosidase ginsenoside biotransformation ginsenoside Rg1 ginsenoside Re |
| url | https://www.mdpi.com/2218-273X/14/12/1611 |
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