Purification and characterization of a novel ginsenoside Rc-hydrolyzing β-glucosidase from Armillaria mellea mycelia

Ginsenosides are the principal compounds responsible for the pharmacological effects and health benefits of Panax ginseng root. Among protopanaxadiol (PPD)-type ginsenosides, minor ginsenosides such as ginsenoside (G)-F2, G-Rh2, compound (C)-Mc1, C-Mc, C-O, C-Y, and C-K are known to be more pharmaco...

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Autores principales: Upadhyaya, Jitendra, Yoon, Min-Sun, Kim, Min-Ji, Ryu, Nam-Soo, Song, Young-Eun, Kim, Young-Hoi, Kim, Myung-Kon
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2016
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5106418/
https://www.ncbi.nlm.nih.gov/pubmed/27837549
http://dx.doi.org/10.1186/s13568-016-0277-x
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author Upadhyaya, Jitendra
Yoon, Min-Sun
Kim, Min-Ji
Ryu, Nam-Soo
Song, Young-Eun
Kim, Young-Hoi
Kim, Myung-Kon
author_facet Upadhyaya, Jitendra
Yoon, Min-Sun
Kim, Min-Ji
Ryu, Nam-Soo
Song, Young-Eun
Kim, Young-Hoi
Kim, Myung-Kon
author_sort Upadhyaya, Jitendra
collection PubMed
description Ginsenosides are the principal compounds responsible for the pharmacological effects and health benefits of Panax ginseng root. Among protopanaxadiol (PPD)-type ginsenosides, minor ginsenosides such as ginsenoside (G)-F2, G-Rh2, compound (C)-Mc1, C-Mc, C-O, C-Y, and C-K are known to be more pharmacologically active constituents than major ginsenosides such as G-Rb1, G-Rb2, G-Rc, and G-Rd. A novel ginsenoside Rc-hydrolyzing β-glucosidase (BG-1) from Armillaria mellea mycelia was purified as a single protein band with molecular weight of 121.5 kDa on SDS-PAGE and a specific activity of 17.9 U mg(−1) protein. BG-1 concurrently hydrolyzed α-(1 → 6)-arabinofuranosidic linkage at the C-20 site or outer β-(1 → 2)-glucosidic linkage at the C-3 site of G-Rc to produce G-Rd and C-Mc1, respectively. The enzyme also hydrolyzed outer and inner glucosidic linkages at the C-3 site of G-Rd to produce C-K via G-F2, and inner glucosidic linkage at the C-3 site of C-Mc1 to produce C-Mc. C-Mc was also slowly hydrolyzed α-(1 → 6)-arabinofuranosidic linkage at the C-20 site to produce C-K with reaction time prolongation. Finally, the pathways for formation of C-Mc and C-K from G-Rc by BG-1 were G-Rc → C-Mc1 → C-Mc and G-Rc → G-Rd → G-F2 → C-K, respectively. The optimum reaction conditions for C-Mc and C-K formation from G-Rc by BG-1 were pH 4.0–4.5, temperature 45–60 °C, and reaction time 72–96 h. This is the first report of efficient production of minor ginsenosides, C-Mc and C-K from G-Rc by β-glucosidase purified from A. mellea mycelia.
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spelling pubmed-51064182016-12-02 Purification and characterization of a novel ginsenoside Rc-hydrolyzing β-glucosidase from Armillaria mellea mycelia Upadhyaya, Jitendra Yoon, Min-Sun Kim, Min-Ji Ryu, Nam-Soo Song, Young-Eun Kim, Young-Hoi Kim, Myung-Kon AMB Express Original Article Ginsenosides are the principal compounds responsible for the pharmacological effects and health benefits of Panax ginseng root. Among protopanaxadiol (PPD)-type ginsenosides, minor ginsenosides such as ginsenoside (G)-F2, G-Rh2, compound (C)-Mc1, C-Mc, C-O, C-Y, and C-K are known to be more pharmacologically active constituents than major ginsenosides such as G-Rb1, G-Rb2, G-Rc, and G-Rd. A novel ginsenoside Rc-hydrolyzing β-glucosidase (BG-1) from Armillaria mellea mycelia was purified as a single protein band with molecular weight of 121.5 kDa on SDS-PAGE and a specific activity of 17.9 U mg(−1) protein. BG-1 concurrently hydrolyzed α-(1 → 6)-arabinofuranosidic linkage at the C-20 site or outer β-(1 → 2)-glucosidic linkage at the C-3 site of G-Rc to produce G-Rd and C-Mc1, respectively. The enzyme also hydrolyzed outer and inner glucosidic linkages at the C-3 site of G-Rd to produce C-K via G-F2, and inner glucosidic linkage at the C-3 site of C-Mc1 to produce C-Mc. C-Mc was also slowly hydrolyzed α-(1 → 6)-arabinofuranosidic linkage at the C-20 site to produce C-K with reaction time prolongation. Finally, the pathways for formation of C-Mc and C-K from G-Rc by BG-1 were G-Rc → C-Mc1 → C-Mc and G-Rc → G-Rd → G-F2 → C-K, respectively. The optimum reaction conditions for C-Mc and C-K formation from G-Rc by BG-1 were pH 4.0–4.5, temperature 45–60 °C, and reaction time 72–96 h. This is the first report of efficient production of minor ginsenosides, C-Mc and C-K from G-Rc by β-glucosidase purified from A. mellea mycelia. Springer Berlin Heidelberg 2016-11-11 /pmc/articles/PMC5106418/ /pubmed/27837549 http://dx.doi.org/10.1186/s13568-016-0277-x Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Original Article
Upadhyaya, Jitendra
Yoon, Min-Sun
Kim, Min-Ji
Ryu, Nam-Soo
Song, Young-Eun
Kim, Young-Hoi
Kim, Myung-Kon
Purification and characterization of a novel ginsenoside Rc-hydrolyzing β-glucosidase from Armillaria mellea mycelia
title Purification and characterization of a novel ginsenoside Rc-hydrolyzing β-glucosidase from Armillaria mellea mycelia
title_full Purification and characterization of a novel ginsenoside Rc-hydrolyzing β-glucosidase from Armillaria mellea mycelia
title_fullStr Purification and characterization of a novel ginsenoside Rc-hydrolyzing β-glucosidase from Armillaria mellea mycelia
title_full_unstemmed Purification and characterization of a novel ginsenoside Rc-hydrolyzing β-glucosidase from Armillaria mellea mycelia
title_short Purification and characterization of a novel ginsenoside Rc-hydrolyzing β-glucosidase from Armillaria mellea mycelia
title_sort purification and characterization of a novel ginsenoside rc-hydrolyzing β-glucosidase from armillaria mellea mycelia
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5106418/
https://www.ncbi.nlm.nih.gov/pubmed/27837549
http://dx.doi.org/10.1186/s13568-016-0277-x
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