Complete Biotransformation of Protopanaxatriol-Type Ginsenosides in Panax ginseng Leaf Extract to Aglycon Protopanaxatriol by β-Glycosidases from Dictyoglomus turgidum and Pyrococcus furiosus
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Aglycon protopanaxatriol (APPT) has valuable pharmacological effects such as memory enhancement and tumor inhibition. β-Glycosidase from the hyperthermophilic bacterium Dictyoglomus turgidum (DT-bgl) hydrolyzes the glucose residues linked to APPT, but not other glycoside residues. β-Glycosidase from the hyperthermophilic bacterium Pyrococcus furiosus (PF-bgl) hydrolyzes the outer sugar at C-6 but not the inner glucose at C-6 or the glucose at C-20. Thus, the combined use of DT-bgl and PF-bgl is expected to increase the biotransformation of PPT-type ginsenosides to APPT. We optimized the ratio of PF-bgl to DTbgl, the concentrations of substrate and enzyme, and the reaction time to increase the biotransformation of ginsenoside Re and PPT-type ginsenosides in Panax ginseng leaf extract to APPT. DT-bgl combined with PF-bgl converted 1.0 mg/ml PPT-type ginsenosides in ginseng leaf extract to 0.58 mg/ml APPT without other ginsenosides, with a molar conversion of 100%. We achieved the complete biotransformation of ginsenoside Re and PPT-type ginsenosides in ginseng leaf extract to APPT by the combined use of two β-glycosidases, suggesting that discarded ginseng leaves can be used as a source of the valuable ginsenoside APPT. To the best of our knowledge, this is the first quantitative production of APPT using ginsenoside Re, and we report the highest concentration and productivity of APPT from ginseng extract to date.
Abstract
Aglycon protopanaxatriol (APPT) has valuable pharmacological effects such as memory enhancement and tumor inhibition. β-Glycosidase from the hyperthermophilic bacterium Dictyoglomus turgidum (DT-bgl) hydrolyzes the glucose residues linked to APPT, but not other glycoside residues. β-Glycosidase from the hyperthermophilic bacterium Pyrococcus furiosus (PF-bgl) hydrolyzes the outer sugar at C-6 but not the inner glucose at C-6 or the glucose at C-20. Thus, the combined use of DT-bgl and PF-bgl is expected to increase the biotransformation of PPT-type ginsenosides to APPT. We optimized the ratio of PF-bgl to DTbgl, the concentrations of substrate and enzyme, and the reaction time to increase the biotransformation of ginsenoside Re and PPT-type ginsenosides in Panax ginseng leaf extract to APPT. DT-bgl combined with PF-bgl converted 1.0 mg/ml PPT-type ginsenosides in ginseng leaf extract to 0.58 mg/ml APPT without other ginsenosides, with a molar conversion of 100%. We achieved the complete biotransformation of ginsenoside Re and PPT-type ginsenosides in ginseng leaf extract to APPT by the combined use of two β-glycosidases, suggesting that discarded ginseng leaves can be used as a source of the valuable ginsenoside APPT. To the best of our knowledge, this is the first quantitative production of APPT using ginsenoside Re, and we report the highest concentration and productivity of APPT from ginseng extract to date.
참고문헌 (21)
- Shin KC, "Classification of glycosidases that hydrolyze the specific positions and types of sugar moieties in ginsenosides", Crit. Rev. Biotechnol., 36, pp.1036~1049 (2016)[DOI]
- Xie JT, "Anti-diabetic effect of ginsenoside Re in ob", ob mice, 2005, pp.1740, 319~325
- Cheng Y, "Anti-amnestic and antiaging effects of ginsenoside Rg1 and Rb1 and its mechanism of action", Acta Pharmacol. Sin., 26, pp.143~149 (2005)[DOI]
- 김희진, "A comprehensive review of the therapeutic and pharmacological effects of ginseng and ginsenosides in central nervous system", Journal of Ginseng Research, 37(1), pp.8~29 (2013), 고려인삼학회
- Naijing Li, "A UPLC", MS-based metabolomics investigation of the protective effect of ginsenosides Rg1 and Rg2 in mice with Alzheimer’s disease, 2016, pp.40(1), 9~17[DOI]
- Lee EH, "Ginsenoside F1 protects human HaCaT keratinocytes from ultraviolet-B-induced apoptosis by maintaining constant levels of Bcl-2", J. Invest. Dermatol., 121, pp.607~613 (2003)[DOI]
- Jeong JJ, "Anti-inflammatory effects of vina-ginsenoside R2 and majonoside R2 isolated from Panax vietnamensis and their metabolites in lipopolysaccharide-stimulated macrophages", Int. Immunopharmacol., 28, pp.700~706 (2015)[DOI]
- Myung Kyum Kim, "Microbial Conversion of Major Ginsenoside Rb1 to Pharmaceutically Active Minor Ginsenoside Rd", The Journal of Microbiology, 43(5), pp.456~462 (2005), 한국미생물학회
- Bae EA, "Metabolism of ginsenoside Re by human intestinal microflora and its estrogenic effect", Biol. Pharm. Bull., 28, pp.1903~1908 (2005)[DOI]
- Wang YZ, "Improvement of memory in mice and increase of hippocampal excitability in rats by ginsenoside Rg1's metabolites ginsenoside Rh1 and protopanaxatriol", J. Pharmacol. Sci., 109, pp.504~510 (2009)[DOI]
- Hasegawa H, "Prevention of growth and metastasis of murine melanoma through enhanced natural-killer cytotoxicity by fatty acid-conjugate of protopanaxatriol", Biol. Pharm. Bull., 25, pp.861~866 (2002)
- Cui CH, "Characterization of the ginsenoside-transforming recombinant beta-glucosidase from Actinosynnema mirum and bioconversion of major ginsenosides into minor ginsenosides", Appl. Microbiol. Biotechnol., 97, pp.649~659 (2013)[DOI]
- Liu L, "Microbial conversion of rare ginsenoside Rf to 20(S)-protopanaxatriol by Aspergillus niger", Biosci. Biotechnol. Biochem., 74, pp.96~100 (2010)[DOI]
- Lee GW, "Beta-glucosidase from Penicillium aculeatum hydrolyzes exo-, 3-O-, and 6-O-beta-glucosides but not 20-O-beta-glucoside and other glycosides of ginsenosides", Appl. Microbiol. Biotechnol., 97, pp.6315~6324 (2013)
- Wang DM, "Enzyme kinetics of ginsenosidase type IV hydrolyzing 6-O-multiglycosides of protopanaxatriol type ginsenosides", Process Biochem., 47, pp.133~138 (2012)[DOI]
- Lee HJ, "Production of aglycone protopanaxatriol from ginseng root extract using Dictyoglomus turgidum beta-glycosidase that specifically hydrolyzes the xylose at the C-6 position and the glucose in protopanaxatriol-type ginsenosides", Appl. Microbiol. Biotechnol., 98, pp.3659~3667 (2014)[DOI]
- Jiang Ning Hu, "Determination of Ginsenosides Content in Korean Ginseng Seeds and Roots by High Performance Liquid Chromatography", Food Science and Biotechnology, 17(2), pp.430~433 (2008), 한국식품과학회
- Shi W, "Investigation of ginsenosides in different parts and ages of Panax ginseng", Food Chem., 102, pp.664~668 (2007)[DOI]
- Yahara S, "Saponins of buds and flowers of Panax ginseng CA Meyer. (1). Isolation of ginsenosides Rd, Re, and Rg1", Chem. Pharm. Bull., 24, pp.3212~3213 (1976)[DOI]
- Shin KC, "Substrate specificity of betaglucosidase from Gordonia terrae for ginsenosides and its application in the production of ginsenosides Rg3, Rg2, and Rh1 from ginseng root extract", J. Biosci. Bioeng., 119, pp.497~504 (2015)
- Oh HJ, "Production of ginsenosides Rg1 and Rh1 by hydrolyzing the outer glycoside at the C-6 position in protopanaxatriol-type ginsenosides using <TEX>${\beta}$<", TEX>-glucosidase from Pyrococcus furiosus, 2014, pp.36, 113~119
