Journal of Ginseng Research

The Korean Society of Ginseng (고려인삼학회)
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Glycosyltransformation of ginsenoside Rh2 into two novel ginsenosides using recombinant glycosyltransferase from Lactobacillus rhamnosus and its in vitro applications

  • Wang, Dan-Dan (School of Life Sciences, Yantai University) ;
  • Kim, Yeon-Ju (Department of Oriental Medicinal Biotechnology, Ginseng Bank, College of Life Science, Kyung Hee University) ;
  • Baek, Nam In (Department of Oriental Medicinal Biotechnology, Ginseng Bank, College of Life Science, Kyung Hee University) ;
  • Mathiyalagan, Ramya (Graduate School of Biotechnology, College of Life Science, Kyung Hee University) ;
  • Wang, Chao (Department of Oriental Medicinal Biotechnology, Ginseng Bank, College of Life Science, Kyung Hee University) ;
  • Jin, Yan (Department of Oriental Medicinal Biotechnology, Ginseng Bank, College of Life Science, Kyung Hee University) ;
  • Xu, Xing Yue (Graduate School of Biotechnology, College of Life Science, Kyung Hee University) ;
  • Yang, Deok-Chun (Department of Oriental Medicinal Biotechnology, Ginseng Bank, College of Life Science, Kyung Hee University)
  • Received : 2018.07.02
  • Accepted : 2019.11.01
  • Published : 2021.01.15
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Abstract

Background: Ginsenoside Rh2 is well known for many pharmacological activities, such as anticancer, antidiabetes, antiinflammatory, and antiobesity properties. Glycosyltransferases (GTs) are ubiquitous enzymes present in nature and are widely used for the synthesis of oligosaccharides, polysaccharides, glycoconjugates, and novel derivatives. We aimed to synthesize new ginsenosides from Rh2 using the recombinant GT enzyme and investigate its cytotoxicity with diverse cell lines. Methods: We have used a GT gene with 1,224-bp gene sequence cloned from Lactobacillus rhamnosus (LRGT) and then expressed in Escherichia coli BL21 (DE3). The recombinant GT protein was purified and demonstrated to transform Rh2 into two novel ginsenosides, and they were characterized by nuclear magnetic resonance (NMR) techniques and evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide assay. Results: Two novel ginsenosides with an additional glucopyranosyl (6→1) and two additional glucopyranosyl (6→1) linked with the C-3 position of the substrate Rh2 were synthesized, respectively. Cell viability assay in the lung cancer (A549) cell line showed that glucosyl ginsenoside Rh2 inhibited cell viability more potently than ginsenoside Rg3 and Rh2 at a concentration of 10 μM. Furthermore, glucosyl ginsenoside Rh2 did not exhibit any cytotoxic effect in murine macrophage cells (RAW264.7), mouse embryo fibroblasts cells (3T3-L1), and skin cells (B16BL6) at a concentration of 10 μM compared with ginsenoside Rh2 and Rg3. Conclusion: This is the first report on the synthesis of two novel ginsenosides, namely, glucosyl ginsenoside Rh2 and diglucosyl ginsenoside Rh2 from Rh2 by using recombinant GT isolated from L. rhamnosus. Moreover, diglucosyl ginsenoside Rh2 might be a new candidate for treatment of inflammation, obesity, and skin whiting, and especially for anticancer.

Keywords

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