Biotechnology and Bioprocess Engineering 2021; 26(6): 968-975  
Biotransformation of Ginsenoside Rb1 to Ginsenoside F2 by Recombinant β-glucosidase from Rat Intestinal Enterococcus gallinarum
Chunlong Yan, Chunyun Hao, Wanzhu Jin, Wei-Wei Dong, and Lin-Hu Quan
Chunlong Yan, Wei-Wei Dong*
Agriculture College of Yanbian University, Yanji, Jilin 133002, China
E-mai: wwdong@ybu.edu.cn
Chunyun Hao
College of Integration Science, Yanbian University, Yanji, Jilin 133002, China
Wanzhu Jin
Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China
University of Chinese Academy of Sciences, Beijing 100049, China
Lin-Hu Quan*
College of Pharmacy, Yanbian University, Yanji, Jilin 133002, China
Tel: +86-433-2436452; Fax: 86-433-2432456
E-mail: lhquan@ybu.edu.cn
These authors contributed equally to this work.
Received: January 6, 2021; Revised: March 17, 2021; Accepted: March 17, 2021; Published online: December 31, 2021.
© The Korean Society for Biotechnology and Bioengineering. All rights reserved.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Panax ginseng is used as a traditional medicine and functional food in several Asian countries. Intestinal bacteria play an important role in the metabolism of components and the production of bioactive metabolites following oral consumption of P. ginseng. We researched the genetic and biochemical properties of the gene encoding β-glucosidase of rat intestinal bacteria that carries out the hydrolytic metabolic reactions. We isolated the ginsenoside transforming Enterococcus gallinarum GM2 from rat colonic contents. After cloning the GH family 3 domain protein β-glucosidase gene (Bgy3) from E. gallinarum, we expressed and purified recombinant Bgy3 protein and then characterized it. We used LC-MS/MS to determine the metabolic profile of ginsenoside Rb1 generated by Bgy3. At pH 7.0 and 40°C, Bgy3 selectively removed the outer sugars of C-20 and C-3, and it produced ginsenoside F2 from ginsenoside Rb1, with a corresponding molar conversion yield of 45%. Bgy3 was found to hydrolyze the ginsenoside Rb1 according to the following pathways: Rb1 → gypenoside XVII → F2, or Rb1 → Rd → F2. Bgy3 functions in the metabolism of ginsenoside Rb1, effectively converting it into ginsenoside F2. This study clearly elucidated the relationship between intestinal bacteria and the metabolism of ginsenoside.
Keywords: ginsenosode Rb1, intestinal bacteria, Enterococcus gallinarum, β-glucosidase, metabolite


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