Gram-Scale Production of Ginsenoside F1 Using a Recombinant Bacterial β-Glucosidase
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초록
Naturally occurring ginsenoside F1 (20-O-β-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 β-glucosidase (BgpA) cloned from a soil bacterium, Terrabacter ginsenosidimutans Gsoil 3082T. 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 gramscale F1 production via recombinant enzyme-mediated biotransformation.
Abstract
Naturally occurring ginsenoside F1 (20-O-β-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 β-glucosidase (BgpA) cloned from a soil bacterium, Terrabacter ginsenosidimutans Gsoil 3082T. 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 gramscale F1 production via recombinant enzyme-mediated biotransformation.
참고문헌 (24)
- Minkyung Bae, "Protective effect of Korean Red Ginseng extract against Helicobacter pylori-induced gastric inflammation in Mongolian gerbils", Journal of Ginseng Research, 38(1), pp.8~15 (2014), 고려인삼학회[DOI]
- 이창호, "A review on the medicinal potentials of ginseng and ginsenosides on cardiovascular diseases", Journal of Ginseng Research, 38(3), pp.161~166 (2014), 고려인삼학회
- Ho Jae Park, "Ginseng in the traditional herbal prescriptions", Journal of Ginseng Research, 36(3), pp.225~241 (2012), 고려인삼학회
- Christensen LP., "Ginsenosides. Chemistry, biosynthesis, analysis, and potential health effects", Adv. Food Nutr. Res, 55, pp.1~99 (2008)
- Corbit RM, "Simplified extraction of ginsenosides from American ginseng (Panax quinquefolius L.) for high-performance liquid chromatographyultraviolet analysis", J. Agric. Food Chem, 53, pp.9867~9873 (2005)
- Nguyen HT, "Dammarane-type saponins from the flower buds of Panax ginseng and their effects on human leukemia cells", Bioorg. Med. Chem. Lett, 20, pp.309~314 (2010)
- Qu C, "Study on ginsenosides in different parts and ages of Panax quinquefolius L", Food Chem, 115, pp.340~346 (2009)
- Shi W, "Investigation of ginsenosides in different parts and ages of Panax ginseng", Food Chem, 102, pp.664~668 (2007)
- Hasegawa H., "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, pp.153~157 (2004)
- Akao T, "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, pp.245~249 (1998)
- Liu Y, "Ginsenoside metabolites, rather than naturally occurring ginsenosides, lead to inhibition of human cytochrome P450enzymes", Toxicol. Sci, 91, pp.356~364 (2006)
- Baek NI, "Cytotoxicities of ginseng saponins and their degradation products against some cancer cell lines", Arch. Pharm. Res, 18, pp.164~168 (1995)
- Popovich DG, "Generation of ginsenosides Rg3and Rh2 from North American ginseng", Phytochemistry, 65, pp.337~344 (2004)
- Park CS, "Biotransformation of ginsenosides by hydrolyzing the sugar moieties of ginsenosides using microbial glycosidases", Appl. Microbiol. Biotechnol, 87, pp.9~19 (2010)
- Ioannis Georgiadis, "Beneficial Health Effects of Chios Gum Mastic and Peroxisome Proliferator-Activated Receptors: Indications of Common Mechanisms", Journal of Medicinal Food, 18(1), pp.1~10 (2015), 한국식품영양과학회[DOI]
- Yu Wang, "Production of ginsenoside F1 using commercial enzyme Cellulase KN", Journal of Ginseng Research, 40(2), pp.121~126 (2016), 고려인삼학회
- An DS, "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, pp.5827~5836 (2010)
- Dae Sung Yoo, "Ginsenoside F1 Modulates Cellular Responses of Skin Melanoma Cells", Journal of Ginseng Research, 35(1), pp.86~91 (2011), 고려인삼학회
- 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)
- Ko SR, "Enzymatic preparation of ginsenosides Rg2, Rh1, and F1", Chem. Pharm. Bull, 51, pp.404~408 (2003)
- Kim YS, "Ginsenoside F1 production from ginsenoside Rg1 by a purified β-glucosidase from Fusarium moniliforme var. subglutinans", Biotechnol. Lett, 33, pp.2457~2461 (2011)
- Wang D, "Enzyme kinetics of ginsenosidase type IV hydrolyzing 6-O-multi-glycosides of protopanaxatriol type ginsenosides", Process Biochem, 47, pp.133~138 (2012)
- Wei Y, "Purification and characterization of a novel and unique ginsenoside Rg 1-hydrolyzing β-D-glucosidase from Penicillium sclerotiorum", Acta Biochim. Biophys. Sin, 43, pp.226~231 (2011)
- An DS, "Characterization of a novel ginsenoside-hydrolyzing α-L-arabinofuranosidase, AbfA, from Rhodanobacter ginsenosidimutans Gsoil 3054T", Appl. Microbiol. Biotechnol, 94, pp.673~682 (2012)