Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Gram-Scale Production of Ginsenoside F1 Using a Recombinant Bacterial β-Glucosidase

  • An, Dong-Shan (Biological Resource Center/Korean Collection for Type Cultures (KCTC), Korea Research Institute of Bioscience and Biotechnology) ;
  • Cui, Chang-Hao (KAIST Institute for Biocentury, Korea Advanced Institute of Science and Technology) ;
  • Siddiqi, Muhammad Zubair (Department of Biotechnology, Hankyong National University) ;
  • Yu, Hong Shan (College of Biotechnology, Dalian Polytechnic University) ;
  • Jin, Feng-Xie (College of Biotechnology, Dalian Polytechnic University) ;
  • Kim, Song-Gun (Biological Resource Center/Korean Collection for Type Cultures (KCTC), Korea Research Institute of Bioscience and Biotechnology) ;
  • Im, Wan-Taek (Department of Biotechnology, Hankyong National University)
  • Received : 2017.03.06
  • Accepted : 2017.06.19
  • Published : 2017.09.28
Download PDF
⟨ Previous Next ⟩

Abstract

Naturally occurring ginsenoside F1 (20-O-${\beta}$-$\text\tiny{D}$-glucopyranosyl-20(S)-protopanaxatriol) is rare. Here, we produced gram-scale quantities of ginsenoside F1 from a crude protopanaxatriol saponin mixture comprised mainly of Re and Rg1 through enzyme-mediated biotransformation using recombinant ${\beta}$-glucosidase (BgpA) cloned from a soil bacterium, Terrabacter ginsenosidimutans Gsoil $3082^T$. In a systematic step-by-step process, the concentrations of substrate, enzyme, and NaCl were determined for maximal production of F1. At an optimized NaCl concentration of 200 mM, the protopanaxatriol saponin mixture (25 mg/ml) was incubated with recombinant BgpA (20 mg/ml) for 3 days in a 2.4 L reaction. Following octadecylsilyl silica gel column chromatography, 9.6 g of F1 was obtained from 60 g of substrate mixture at 95% purity, as assessed by chromatography. These results represent the first report of gram-scale F1 production via recombinant enzyme-mediated biotransformation.

Keywords

References

  1. Bae M, Jang S, Lim JW, Kang J, Bak EJ, Cha JH, et al. 2014. Protective effect of Korean Red Ginseng extract against Helicobacter pylori-induced gastric inflammation in Mongolian gerbils. J. Ginseng Res. 38: 8-15. https://doi.org/10.1016/j.jgr.2013.11.005
  2. Lee CH, Kim JH. 2014. A review on the medicinal potentials of ginseng and ginsenosides on cardiovascular diseases. J. Ginseng Res. 38: 161-166. https://doi.org/10.1016/j.jgr.2014.03.001
  3. Park HJ, Kim DH, Park SJ, Kim JM, Ryu JH. 2012. Ginseng in traditional herbal prescriptions. J. Ginseng Res. 36: 225-241. https://doi.org/10.5142/jgr.2012.36.3.225
  4. Christensen LP. 2008. Ginsenosides. Chemistry, biosynthesis, analysis, and potential health effects. Adv. Food Nutr. Res. 55: 1-99.
  5. Corbit RM, Ferreira JF, Ebbs SD, Murphy LL. 2005. Simplified extraction of ginsenosides from American ginseng (Panax quinquefolius L.) for high-performance liquid chromatographyultraviolet analysis. J. Agric. Food Chem. 53: 9867-9873. https://doi.org/10.1021/jf051504p
  6. Nguyen H T, S ong GY, Kim JA, H yun JH, Kang H K, K im YH. 2010. D ammarane-type saponins f rom the f lower buds of Panax ginseng and their effects on human leukemia cells. Bioorg. Med. Chem. Lett. 20: 309-314. https://doi.org/10.1016/j.bmcl.2009.10.110
  7. Qu C, Bai Y, Jin X, Wang Y, Zhang K, You J, et al. 2009. Study on ginsenosides in different parts and ages of Panax quinquefolius L. Food Chem. 115: 340-346. https://doi.org/10.1016/j.foodchem.2008.11.079
  8. Shi W, Wang Y, Li J, Zhang H, Ding L. 2007. Investigation of ginsenosides in different parts and ages of Panax ginseng. Food Chem. 102: 664-668. https://doi.org/10.1016/j.foodchem.2006.05.053
  9. Hasegawa H. 2004. Proof of the mysterious efficacy of ginseng: basic and clinical trials: metabolic activation of ginsenoside: deglycosylation by intestinal bacteria and esterification with fatty acid. J. Pharmacol. Sci. 95: 153-157. https://doi.org/10.1254/jphs.FMJ04001X4
  10. Akao T, Kanaoka M, Kobashi K. 1998. Appearance of compound K, a major metabolite of ginsenoside Rb1 by intestinal bacteria, in rat plasma after oral administration - measurement of compound K by enzyme immunoassay. Biol. Pharm. Bull. 21: 245-249. https://doi.org/10.1248/bpb.21.245
  11. Liu Y, Zhang JW, Li W, Ma H, Sun J, Deng MC, Yang L. 2006. Ginsenoside metabolites, rather than naturally occurring ginsenosides, lead to inhibition of human cytochrome P450 enzymes. Toxicol. Sci. 91: 356-364. https://doi.org/10.1093/toxsci/kfj164
  12. Baek NI, Kim DS, Lee YH, Park JD, Lee CB, Kim SI. 1995. Cytotoxicities of ginseng saponins and their degradation products against some cancer cell lines. Arch. Pharm. Res. 18: 164-168. https://doi.org/10.1007/BF02979189
  13. Popovich DG, Kitts DD. 2004. Generation of ginsenosides Rg3 and Rh2 from North American ginseng. Phytochemistry 65: 337-344. https://doi.org/10.1016/j.phytochem.2003.11.020
  14. Park CS, Yoo MH, Noh KH, Oh DK. 2010. Biotransformation of ginsenosides by hydrolyzing the sugar moieties of ginsenosides using microbial glycosidases. Appl. Microbiol. Biotechnol. 87: 9-19. https://doi.org/10.1007/s00253-010-2567-6
  15. Siddiqi MZ, Siddiqi MH, Kim YJ, J in Y, Huq MA, Yang DC. 2015. Effect of fermented red ginseng extract enriched in ginsenoside Rg3 on the differentiation and mineralization of preosteoblastic MC3T3-E1 cells. J. Med. Food 18: 1-7. https://doi.org/10.1089/jmf.2014.0021
  16. Wang Y, Choi KD, Yu H, Jin F, Im WT. 2016. Production of ginsenoside F1 using commercial enzyme Cellulase KN. J. Ginseng Res. 40: 121-126. https://doi.org/10.1016/j.jgr.2015.06.003
  17. An DS, Cui CH, Lee HG, Wang L, Kim SC, Lee ST, et al. 2010. Identification and characterization of a novel Terrabacter ginsenosidimutans sp. nov. beta-glucosidase that transforms ginsenoside Rb1 into the rare gypenosides XVII and LXXV. Appl. Environ. Microbiol. 76: 5827-5836. https://doi.org/10.1128/AEM.00106-10
  18. Yoo DS, Rho HS, Lee YG, Yeom MH, Kim DH, Lee SL, et al. 2011. Ginsenoside F1 modulates cellular responses of skin Melanoma Cells. J. Ginseng Res. 35: 86-91. https://doi.org/10.5142/jgr.2011.35.1.086
  19. Lee EH, Cho SY, Kim SJ, Shin ES, Chang HK, Kim DH, et al. 2003. Ginsenoside F1 protects human HaCaT keratinocytes from ultraviolet-B-induced apoptosis by maintaining constant levels of Bcl-2. J. Invest. Dermatol. 121: 607-613. https://doi.org/10.1046/j.1523-1747.2003.12425.x
  20. Ko SR, Choi KJ, Suzuki K, Suzuki Y. 2003. Enzymatic preparation of ginsenosides Rg2, Rh1, and F1. Chem. Pharm. Bull. 51: 404-408. https://doi.org/10.1248/cpb.51.404
  21. Kim YS, Yoo MH, Lee GW, Choi JG, Kim KR, Oh DK. 2011. Ginsenoside F1 production from ginsenoside Rg1 by a purified $\beta$-glucosidase from Fusarium moniliforme var. subglutinans. Biotechnol. Lett. 33: 2457-2461. https://doi.org/10.1007/s10529-011-0719-0
  22. Wang D, Yu H, Song J, Xu Y, Jin F. 2012. Enzyme kinetics of ginsenosidase type IV hydrolyzing 6-O-multi-glycosides of protopanaxatriol type ginsenosides. Process Biochem. 47: 133-138. https://doi.org/10.1016/j.procbio.2011.10.026
  23. Wei Y, Zhao W, Zhang Q, Zhao Y, Zhang Y. 2011. Purification and characterization of a novel and unique ginsenoside Rg 1-hydrolyzing $\beta$-D-glucosidase from Penicillium sclerotiorum. Acta Biochim. Biophys. Sin. 43: 226-231. https://doi.org/10.1093/abbs/gmr001
  24. An DS, Cui CH, Sung BH, Yang HC, Kim SC, Lee ST, et al. 2012. Characterization of a novel ginsenoside-hydrolyzing $\alpha$-L-arabinofuranosidase, AbfA, from Rhodanobacter ginsenosidimutans Gsoil 3054T. Appl. Microbiol. Biotechnol. 94: 673-682. https://doi.org/10.1007/s00253-011-3614-7

Cited by

  1. Pro-angiogenic Ginsenosides F1 and Rh1 Inhibit Vascular Leakage by Modulating NR4A1 vol.9, pp.None, 2019, https://doi.org/10.1038/s41598-019-41115-2
  2. Gypenoside LXXV Promotes Cutaneous Wound Healing In Vivo by Enhancing Connective Tissue Growth Factor Levels Via the Glucocorticoid Receptor Pathway vol.24, pp.8, 2017, https://doi.org/10.3390/molecules24081595
  3. Enhanced Production of Protopanaxatriol from Ginsenoside Re and Rg1 Using a Recombinant Bacterial β-glucosidase vol.24, pp.4, 2017, https://doi.org/10.1007/s12257-019-0090-x
  4. High-density immobilization of a ginsenoside-transforming β-glucosidase for enhanced food-grade production of minor ginsenosides vol.103, pp.17, 2017, https://doi.org/10.1007/s00253-019-09951-4
  5. Exploration and Characterization of Novel Glycoside Hydrolases from the Whole Genome of Lactobacillus ginsenosidimutans and Enriched Production of Minor Ginsenoside Rg3( S ) by a Recombinant Enzymat vol.10, pp.2, 2017, https://doi.org/10.3390/biom10020288
  6. Characterization of a Novel Ginsenoside MT1 Produced by an Enzymatic Transrhamnosylation of Protopanaxatriol-Type Ginsenosides Re vol.10, pp.4, 2017, https://doi.org/10.3390/biom10040525
  7. Cumulative Production of Bioactive Rg3, Rg5, Rk1, and CK from Fermented Black Ginseng Using Novel Aspergillus niger KHNT-1 Strain Isolated from Korean Traditional Food vol.9, pp.2, 2017, https://doi.org/10.3390/pr9020227
  8. Chryseobacterium panacisoli sp. nov., isolated from ginseng-cultivation soil with ginsenoside-converting activity vol.71, pp.11, 2017, https://doi.org/10.1099/ijsem.0.005086