• Journal of Ginseng Research
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• v.39 no.3
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• pp.221-229
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• 2015

Background: Minor ginsenosides, those having low content in ginseng, have higher pharmacological activities. To obtain minor ginsenosides, the biotransformation of American ginseng protopanaxadiol (PPD)-ginsenoside was studied using special ginsenosidase type-I from Aspergillus niger g.848. Methods: DEAE (diethylaminoethyl)-cellulose and polyacrylamide gel electrophoresis were used in enzyme purification, thin-layer chromatography and high performance liquid chromatography (HPLC) were used in enzyme hydrolysis and kinetics; crude enzyme was used in minor ginsenoside preparation from PPD-ginsenoside; the products were separated with silica-gel-column, and recognized by HPLC and NMR (Nuclear Magnetic Resonance). Results: The enzyme molecular weight was 75 kDa; the enzyme firstly hydrolyzed the C-20 position 20-O-${\beta}$-D-Glc of ginsenoside Rb1, then the C-3 position 3-O-${\beta}$-D-Glc with the pathway $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}C-K$. However, the enzyme firstly hydrolyzed C-3 position 3-O-${\beta}$-D-Glc of ginsenoside Rb2 and Rc, finally hydrolyzed 20-O-L-Ara with the pathway $Rb2{\rightarrow}C-O{\rightarrow}C-Y{\rightarrow}C-K$, and $Rc{\rightarrow}C-Mc1{\rightarrow}C-Mc{\rightarrow}C-K$. According to enzyme kinetics, $K_m$ and $V_{max}$ of Michaelis-Menten equation, the enzyme reaction velocities on ginsenosides were Rb1 > Rb2 > Rc > Rd. However, the pure enzyme yield was only 3.1%, so crude enzyme was used for minor ginsenoside preparation. When the crude enzyme was reacted in 3% American ginseng PPD-ginsenoside (containing Rb1, Rb2, Rc, and Rd) at $45^{\circ}C$ and pH 5.0 for 18 h, the main products were minor ginsenosides C-Mc, C-Y, F2, and C-K; average molar yields were 43.7% for C-Mc from Rc, 42.4% for C-Y from Rb2, and 69.5% for F2 and C-K from Rb1 and Rd. Conclusion: Four monomer minor ginsenosides were successfully produced (at low-cost) from the PPD-ginsenosides using crude enzyme.

• Proceedings of the Ginseng society Conference
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• 1993.09a
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• pp.74-83
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• 1993

In an attempt toward the synthesis of the difficulty accessible ginseng saponins the four dammarane glycosides identical to the natural $ginsenosides-Rh_2,$ - F2, compound K and chikusetsusaponin - LT8 have been prepared from betulafolienetriol(=dammar-24-ene-$3{\alpha},12{\beta}\;20(S)-triol).\;3-O-{\beta}-D-Glucopyranoside$ of 20(S) - protopanaxadiol $(=ginsenoside-Rh_2)$ have been obtained by the regio - and stereoselective glycosylation of the $12-O-acetyldammar-24-ene-3{\beta},\;12{\beta},$ 20(S)-triol. The 12-ketoderivative of 20(S)-protopanaxadiol has been used as aglycon in synthesis of chikusetsusaponin - LT8. Attempted regio - and stereoselective glycosylation of the less reactive tertiary C - 20 - hydroxyl group in order to synthesize the $20-O-{\beta}-D-glucopyranoside$ of 20(S)-protopanaxadiol(=compound K) using 3, 12 - di - O - acetyldammar - 24 - ene - $3{\beta},12{\beta},20(S)$-trial as aglycon was unsuccessful. Glycosylation of 3, 12 - diketone of betulafolienetriol followed by $NaBH_4$ reduction yielded the $20-O-{\beta}-D-glucopyranoside\;of\;dammar-24-ene-3{\beta},12{\alpha},$ 20(S)-triol, the $12{\alpha}-epimer$ of 20(S) - protopanaxadiol. Moreover, a number of semisynthetic ocotillol - type glucosides, analogs of natural pseudoginsenosides, have been prepared.

• Journal of Ginseng Research
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• v.40 no.4
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• pp.366-374
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• 2016

Background: In this study, we screened and identified an endophyte JG09 having strong biocatalytic activity for ginsenosides from Platycodon grandiflorum, converted ginseng total saponins and ginsenoside monomers, determined the source of minor ginsenosides and the transformation pathways, and calculated the maximum production of minor ginsenosides for the conversion of ginsenoside Rb1 to assess the transformation activity of endophyte JG09. Methods: The transformation of ginseng total saponins and ginsenoside monomers Rb1, Rb2, Rc, Rd, Rg1 into minor ginsenosides F2, C-K and Rh1 using endophyte JG09 isolated by an organizational separation method and Esculin-R2A agar assay, as well as the identification of transformed products via TLC and HPLC, were evaluated. Endophyte JG09 was identified through DNA sequencing and phylogenetic analysis. Results: A total of 32 ${\beta}$-glucosidase-producing endophytes were screened out among the isolated 69 endophytes from P. grandiflorum. An endophyte bacteria JG09 identified as Luteibacter sp. effectively converted protopanaxadiol-type ginsenosides Rb1, Rb2, Rc, Rd into minor ginsenosides F2 and C-K, and converted protopanaxatriol-type ginsenoside Rg1 into minor ginsenoside Rh1. The transformation pathways of major ginsenosides by endophyte JG09 were as follows: $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}C-K$; $Rb2{\rightarrow}C-O{\rightarrow}C-Y{\rightarrow}C-K$; $Rc{\rightarrow}C-Mc1{\rightarrow}C-Mc{\rightarrow}C-K$; $Rg1{\rightarrow}Rh1$. The maximum production rate of ginsenosides F2 and C-K reached 94.53% and 66.34%, respectively. Conclusion: This is the first report about conversion of major ginsenosides into minor ginsenosides by fermentation with P. grandiflorum endophytes. The results of the study indicate endophyte JG09 would be a potential microbial source for obtaining minor ginsenosides.

• Korean Journal of Pharmacognosy
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• v.37 no.4 s.147
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• pp.217-220
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• 2006

This study compared the contents of ginsenoside according to the extract conditions of red ginseng to provide basic information for developing functional food using red ginseng. According to the result, the content of crude saponin was highest in 72 hours of extraction at $82^{\circ}C$ (RG-823). The content of prosapogenin (ginsenoside $Rh_1,\;Rh_2,\;Rg_2,\;Rg_3$) was highest in 48 hours of extraction, and followed by 72 and 24 hours at $82^{\circ}C$. And at $93^{\circ}C$ the prosapogenin contents were highest in the order of 48 hours, and next in 24 and 72 hours. In addition, ginsenoside $Rb_1,\;Rb_2$ Rc and Re were not detected in 72 hours of extraction at $93^{\circ}C$ (RG-933) presumedly due to hydrolysis, but ginsenoside Rd, Rf and $Rg_1$ were detected as long as 72 hours of extraction. These results show that protopanaxatriol group is relatively more resistant to heat than protopanaxadiol group.

• Preventive Nutrition and Food Science
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• v.21 no.4
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• pp.389-392
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• 2016

The present study was conducted to investigate the ginsenoside profiles of the main root, root hair, and leaf of ginseng in order to demonstrate their possible application in medicine. The total ginsenoside content of the leaf was up to 12 times than that in the main root, and the content of protopanaxadiol groups was higher than that of protopanaxatriol groups in all the samples. The leaf was shown to contain high amounts of ginsenosides Rb3 and Rh1, whereas the main root contained large amounts of ginsenosides Rb1 and Rc. Moreover, Rb2, Rb3, and Rg1 were only detected in the root hair, leaf, and main root, respectively. The ginsenoside Re content of Panax ginseng leaf and root hair was 2.6~4 times higher than that of the main root. Therefore, the results indicate that the ginsenoside content of Panax ginseng is higher in the leaf and root hair, and lower in the main root.

• Journal of Microbiology and Biotechnology
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• v.23 no.12
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• pp.1802-1805
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• 2013

Ginsenosides are the most important ingredient of ginseng and are known to possess many pharmacological and biological effects. Rb1, a major protopanaxadiol ginsenoside, is the most abundant ginsenoside in Panax ginseng C.A Meyer and can be hydrolyzed into more pharmaceutically potent minor ginsenosides. To identify a microorganism that is capable of converting Rb1 into other ginsenosides, we screened 12 Microbacterium spp., and M. trichothecenolyticum was identified as a likely candidate. M. trichothecenolyticum converted Rb1 into Rd and then into Rh2 based on TLC and HPLC analyses of reaction products. This biotransformation method can be easily applied for mass production of Rd and Rh2 by using Rb1.

• The Korean Journal of Food And Nutrition
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• v.11 no.1
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• pp.82-87
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• 1998

To study of production of Panax ginseng leaf tea, after harvested the leaves in July, August, and September as ripening season, the content and composition of ginseng saponin were investigated. 1. Crude saponin contents in the leaves were a about 16.5%, and they were found to be lower in the leaf harvested in September than those harvested in July or August. 2. As similar patterns were observed with month to month in ginsenoside, sum of major ginsenosides of -Re, -Rd and -Rg1 was fixed about 70% of saponin at harvested in each month. And minor components were ginsenoside -Rb1, -Rb2 and -Rc as in order. 3. The ratio of protopanaxadiol(PD)/protopanaxatriol(PT) was revealed reduction of 1.13 of harvested in July to 0.85 of those in September gradually. The contents of protopanaxadiol were high in the leaves of August and protopanaxatriol was high in those September.

• Journal of Ginseng Research
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• v.43 no.1
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• pp.116-124
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• 2019

Background: Ginsenosides are known as the principal pharmacological active constituents in Panax medicinal plants such as Asian ginseng, American ginseng, and Notoginseng. Some ginsenosides, especially the 20(R) isomers, are found in trace amounts in natural sources and are difficult to chemically synthesize. The present study provides an approach to produce such trace ginsenosides applying biotransformation through Escherichia coli modified with relevant genes. Methods: Seven uridine diphosphate glycosyltransferase (UGT) genes originating from Panax notoginseng, Medicago sativa, and Bacillus subtilis were synthesized or cloned and constructed into pETM6, an ePathBrick vector, which were then introduced into E. coli BL21star (DE3) separately. 20(R)-Protopanaxadiol (PPD), 20(R)-protopanaxatriol (PPT), and 20(R)-type ginsenosides were used as substrates for biotransformation with recombinant E. coli modified with those UGT genes. Results: E. coli engineered with $GT95^{syn}$ selectively transfers a glucose moiety to the C20 hydroxyl of 20(R)-PPD and 20(R)-PPT to produce 20(R)-CK and 20(R)-F1, respectively. GTK1- and GTC1-modified E. coli glycosylated the C3-OH of 20(R)-PPD to form 20(R)-Rh2. Moreover, E. coli containing $p2GT95^{syn}K1$, a recreated two-step glycosylation pathway via the ePathBrich, implemented the successive glycosylation at C20-OH and C3-OH of 20(R)-PPD and yielded 20(R)-F2 in the biotransformation broth. Conclusion: This study demonstrates that rare 20(R)-ginsenosides can be produced through E. coli engineered with UTG genes.

• Food Science and Biotechnology
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• v.16 no.5
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• pp.848-853
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• 2007

The effects of high-hydrostatic pressure (HHP) on the ginsenoside concentration in Korean red ginseng were investigated. HHP-pretreated Korean red ginseng samples were compared to samples produced by a conventional method. Six-year-old Korean fresh ginseng (Panax ginseng C.A. Meyer) samples were vacuum-packaged in polyethylene film and treated at room temperature for 1 min with HHP (200-600 MPa) and steamed at $98^{\circ}C$ for 3 hr. Major ginsenosides of red ginseng were analyzed by HPLC. HHP-pretreated red ginseng showed a 45% higher level of total major ginsenosides than conventionally prepared red ginseng. The levels of 4 protopanaxadiol-type ginsenosides increased 34-43% and the levels of 5 protopanaxatriol-type ginsenosides increased 45-66%. Scanning electron microscopy and electrical conductivity spectrum analysis showed that HHP pretreatment damaged ginseng plant cells and increased extraction efficiencies of ginsenosides from red ginseng products.

• Journal of Ginseng Research
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• v.22 no.1
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• pp.43-50
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• 1998

We demonstrated in previous study that protopanaxadiol and protopanxatriol saponins show antinociceptive activity in acetic acid induced writhing test and in the second phase (11-40 min) of formalin test but not tail-flick test. To identify further which ginsenoside has antinociceptive activity among various ginseng saponins, we have investigated antinociceptive effects of several ginsenosides using writhing and formalin test. Ginsenoside Rc, Rd, Re, and Rf induced antinociception in writhing test. These four ginsenosides also induced antinociception in the second phase of formalin (11-40 min) test but these ginsenosides showed a slight antinociception in the first phase (010 min) of formalin test except ginsenoside Rf. The antinociceptive effects induced by the ginsenosides were dose dependent and were not blocked by an opioid receptor antagonist, naloxone. The order of antinociceptive potency was Rd > Rc > Re > Rf in the formalin test. However, these ginsenosides did not show any significant analgesic effects in a tail-flick test. These results suggest that ginsenosides such as Rc, Rd, Re, and Rf inhibit tonic pain rather than acute pain induced by noxious heat. These results also indicate that the antinociceptive activity. Induced by ginsenosides may be one of the actions for pharmacological effects of Panax ginseng.