• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.7
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• pp.946-950
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• 2009

Chemical components and antioxidative activities of white ginseng, red ginseng and extruded white ginseng (EWG) were evaluated. Extrusion condition was 20% moisture content, 100 and $140^{\circ}C$ barrel temperature. The results showed that total sugar and acidic polysaccharide contents of white ginseng powder were increased after extrusion treatment of which EWG at $140^{\circ}C$ barrel temperature had higher value than EWG at $100^{\circ}C$ barrel temperature. Free radical scavenging activity of EWG at $140^{\circ}C$ barrel temperature was 80.2 and 45.6% respectively. The butanol fraction of polyphenolic compound and acidic polysaccharide were $27.2{\pm}0.1\;mg/g$ and $217.6{\pm}0.7\;mg/g$, respectively. The ginsenosides were quantified by HPLC and the yield of ginsenoside-Rg3s and Rg3r were achieved by extrusion process.

• Journal of Ginseng Research
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• v.41 no.3
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• pp.392-402
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• 2017

Background: Previously, we reported that Korean Red Ginseng inhibited liver fibrosis in mice and reduced the expressions of fibrogenic genes in hepatic stellate cells (HSCs). The present study was undertaken to identify the major ginsenoside responsible for reducing the numbers of HSCs and the underlying mechanism involved. Methods: Using LX-2 cells (a human immortalized HSC line) and primary activated HSCs, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) assays were conducted to examine the cytotoxic effects of ginsenosides. $H_2O_2$ productions, glutathione contents, lactate dehydrogenase activities, mitochondrial membrane permeabilities, apoptotic cell subpopulations, caspase-3/-7 activities, transferase dUTP nick end labeling (TUNEL) staining, and immunoblot analysis were performed to elucidate the molecular mechanism responsible for ginsenoside-mediated cytotoxicity. Involvement of the AMP-activated protein kinase (AMPK)-related signaling pathway was examined using a chemical inhibitor and small interfering RNA (siRNA) transfection. Results and conclusion: Of the 11 ginsenosides tested, 20S-protopanaxadiol (PPD) showed the most potent cytotoxic activity in both LX-2 cells and primary activated HSCs. Oxidative stress-mediated apoptosis induced by 20S-PPD was blocked by N-acetyl-$\text\tiny L$-cysteine pretreatment. In addition, 20S-PPD concentration-dependently increased the phosphorylation of AMPK, and compound C prevented 20S-PPD-induced cytotoxicity and mitochondrial dysfunction. Moreover, 20S-PPD increased the phosphorylation of liver kinase B1 (LKB1), an upstream kinase of AMPK. Likewise, transfection of LX-2 cells with LKB1 siRNA reduced the cytotoxic effect of 20S-PPD. Thus, 20S-PPD appears to induce HSC apoptosis by activating LKB1-AMPK and to be a therapeutic candidate for the prevention or treatment of liver fibrosis.

• Journal of Ginseng Research
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• v.35 no.2
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• pp.235-242
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• 2011

To obtain microorganisms for the microbial conversion of ginsenosides in red ginseng extract (RGE), mushroom mycelia were used for the fermentation of RGE. After fermentation, total sugar contents and polyohenol contents of the RGEs fermented with various mushrooms were not a significant increase between RGE and the ferments. But uronic acid content was relatively higher in the fermented RGEs cultured with Lentus edodes (2155.6 ${\mu}g/mL$), Phelllinus linteus (1690.9 ${\mu}g/mL$) and Inonotus obliquus 26137 and 26147 (1549.5 and 1670.7 ${\mu}g/mL$) compared to the RGE (1307.1 ${\mu}g/mL$). The RGEs fermented by Ph. linteus, Cordyceps militaris, and Grifola frondosa showed particularly high levels of total ginsenosides (20018.1, 17501.6, and 16267.0 ${\mu}g/mL$, respectively). The ferments with C. militaris (6974.2 ${\mu}g/mL$), Ph. linteus (9109.2 ${\mu}g/mL$), and G. frondosa (7023.0 ${\mu}g/mL$) also showed high levels of metabolites (sum of compound K, $Rh_1$, $Rg_5$, $Rk_1$, $Rg_3$, and $Rg_2$) compared to RGE (3615.9 ${\mu}g/mL$). Among four different RGE concentrations examined, a 20 brix concentration of RGE was favorable for the fermentation of Ph. linteus. Maximum biotransformation of ginsneoside metabolites (9395.5 ${\mu}g/mL$) was obtained after 5 days fermentation with Ph. linteus. Maximum mycelial growth of 2.6 mg/mL was achieved at 9 days, in which growth was not significantly different during 5 to 9 days fermentation. During fermentation of RGE by Ph. linteus in a 7 L fermenter, $Rg_3$, $Rg_5$, and $Rk_1$ contents showed maximum concentrations after 5 days similar to flask fermentation. These results confirm that fermentation with Ph. linteus is very useful for preparing minor ginsenoside metabolites while being safe for foods.

• 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.

• Journal of the East Asian Society of Dietary Life
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• v.23 no.4
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• pp.437-443
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• 2013

Rice wine (makgeolli) containing various amounts of mountain ginsengs (MG) are being prepared with nuruk and yeasts, and the physicochemical characteristics and contents of ginsenosides in MG-makgeolli were analyzed. Average particle size of MG powder is $29.1{\mu}m$. MG slice (20 g) or powder (0~20 g) and rice (3,000 g) were used for 12 days fermentation of makgeolli, makgeolli containing slice of MG (SW-makgeolli), makgeolli containing 2 g (PW1-makgeolli), 10 g (PW2- makgeolli), 20 g (PW3-makgeolli) of powder of MG, respectively. Soluble solids and pH levels show no differences between five kinds of makgeolli groups, whilst the presence of high amounts of MG (PW3-makgeolli) caused decreases in ethanol and acidity. Major free amino acids in MG-makgeolli are glutamic acid and arginine. Total contents of 14 ginsenosides are approximately 2.5 g/100 g of dried MG powder and major ginsenoside were ginsenosides Re, Rb1, Rb2, Rg1, Rc and Rf. During the propagation of makgeolli containing MG, the ginsenosides Rb1, Rb2, Rb3, and Rc decreased, whilst ginsenosides Rg3 and compound K increased highly. It indicates that ginsenosides in MG are metabolized to different forms of ginsenosides by brewing microorganisms.

• Molecules and Cells
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• v.21 no.1
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• pp.52-62
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• 2006

In previous reports we demonstrated that ginsenosides, active ingredients of Panax ginseng, affect some subsets of voltage-dependent $Ca^{2+}$ channels in neuronal cells expressed in Xenopus laevis oocytes. However, the major component(s) of ginseng that affect cloned $Ca^{2+}$ channel subtypes such as ${\alpha}_{1C}$(L)-, ${\alpha}_{1B}$(N)-, ${\alpha}_{1A}$(P/Q)-, ${\alpha}_{1E}$(R)- and ${\alpha}_{1G}$(T) have not been identified. Here, we used the two-microelectrode voltage clamp technique to characterize the effects of ginsenosides and ginsenoside metabolites on $Ba^{2+}$ currents ($I_{Ba}$) in Xenopus oocytes expressing five different $Ca^{2+}$ channel subtypes. Exposure to ginseng total saponins (GTS) induced voltage-dependent, dose-dependent and reversible inhibition of the five channel subtypes, with particularly strong inhibition of the ${\alpha}_{1G}$-type. Of the various ginsenosides, $Rb_1$, Rc, Re, Rf, $Rg_1$, $Rg_3$, and $Rh_2$, ginsenoside $Rg_3$ also inhibited all five channel subtypes and ginsenoside $Rh_2$ had most effect on the ${\alpha}_{1C}$- and ${\alpha}_{1E}$-type $Ca^{2+}$ channels. Compound K (CK), a protopanaxadiol ginsenoside metabolite, strongly inhibited only the ${\alpha}_{1G}$-type of $Ca^{2+}$ channel, whereas M4, a protopanaxatriol ginsenoside metabolite, had almost no effect on any of the channels. $Rg_3$, $Rh_2$, and CK shifted the steady-state activation curves but not the inactivation curves in the depolarizing direction in the ${\alpha}_{1B}$- and ${\alpha}_{1A}$-types. These results reveal that $Rg_3$, $Rh_2$ and CK are the major inhibitors of $Ca^{2+}$ channels in Panax ginseng, and that they show some $Ca^{2+}$ channel selectivity.

• Journal of Pharmacopuncture
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• v.19 no.3
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• pp.213-224
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• 2016

Objectives: Ginseng Rh2+ is enzyme-treated ginseng extract containing high amounts of converted ginsenosides, such as compound k, Rh2, Rg3, which have potent anticancer activity. We conducted general and genetic toxicity tests to evaluate the safety of ginseng Rh2+. Methods: An acute oral toxicity test was performed at a high-level dose of 4,000 mg/kg/day in Sprague-Dawley (SD) rats. A 14-day range-finding study was also conducted to set dose levels for the 90-day study. A subchronic 90-day toxicity study was performed at dose levels of 1,000 and 2,000 mg/kg/day to investigate the no-observed-adverse-effect level (NOAEL) of ginseng Rh2+ and target organs. To identify the mutagenic potential of ginseng Rh2+, we conducted a bacterial reverse mutation test (Ames test) using amino-acid-requiring strains of Salmonella typhimurium and Escherichia coli (E. coli), a chromosome aberration test with Chinese hamster lung (CHL) cells, and an in vivo micronucleus test using ICR mice bone marrow as recommended by the Korean Ministry of Food and Drug Safety. Results: According to the results of the acute oral toxicity study, the approximate lethal dose (ALD) of ginseng Rh2+ was estimated to be higher than 4,000 mg/kg. For the 90-day study, no toxicological effect of ginseng Rh2+ was observed in body-weight changes, food consumption, clinical signs, organ weights, histopathology, ophthalmology, and clinical pathology. The NOAEL of ginseng Rh2+ was established to be 2,000 mg/kg/day, and no target organ was found in this test. In addition, no evidence of mutagenicity was found either on the in vitro genotoxicity tests, including the Ames test and the chromosome aberration test, or on the in vivo in mice bone marrow micronucleus test. Conclusion: On the basis of our findings, ginseng Rh2+ is a non-toxic material with no genotoxicity. We expect that ginseng Rh2+ may be used as a novel adjuvant anticancer agent that is safe for long-term administration.

• Journal of Food Hygiene and Safety
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• v.36 no.5
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• pp.418-429
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• 2021

This paper investigates the antioxidant activity and quality characteristics of yanggaeng containing white ginseng and red ginseng extracts and their enzyme hydrolysates that were produced for the purpose of the study. White and red ginseng extracts were hydrolyzed using Rapidase C80 max, Pyr-flo, and Ultimase MFC. Ginsenoside F2 and compound K (CK) were not detected in white and red ginseng before enzymic reaction but were detected in white and red ginseng hydrolyzed through Rapidase C80 max, Pyr-flo, and Ultimase MFC, and the content of CK was the highest in the second enzymic reaction group of red ginseng. Upon preparing yanggaeng containing white and red ginseng before or after enzymatic hydrolysis, the polyphenol content and antioxidant abilities were analyzed. The yanggaeng containing enzyme-hydrolyzed white ginseng and red ginseng showed greater total polyphenol content, superior DPPH radical scavenging activity, superior ABTS radical scavenging activity, and superior FRAP analysis results compared to the yanggaeng that doesn't contain white or red ginseng. As the enzymic reaction was performed in the added white and red ginseng, the antioxidant activity increased significantly (P<0.05). In brightness(L^*), non-additive yanggaeng (control group) was the highest, red ginseng yanggaeng (RG) showed the highest redness(a^*), and the white ginseng yanggaeng (WG) showed the highest yellowness(b^*). In terms of texture, the yanggaeng containing red ginseng with second hydrolysis (RG-T2) showed significantly high results in hardness, springiness, chewiness, cohesiveness, and gumminess (P<0.05). In conclusion, treating white and red ginseng with Rapidase C80 max, Pyr-flo, and Ultimase MFC is very useful in ginsenoside deglycosylation and will produce CK with excellent biological activity. It can also be seen that yanggaeng containing white and red ginseng hydrolyzed with enzymes significantly increase total polyphenol and antioxidant activity compared to the control group (yanggaeng with no added ginseng). These results will be useful as excellent foundational data for the production of functional yanggaeng in the future.

• Journal of Ginseng Research
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• v.41 no.3
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• pp.419-427
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• 2017

Background: Glycosylation of natural compounds increases the diversity of secondary metabolites. Glycosylation steps are implicated not only in plant growth and development, but also in plant defense responses. Although the activities of uridine-dependent glycosyltransferases (UGTs) have long been recognized, and genes encoding them in several higher plants have been identified, the specific functions of UGTs in planta remain largely unknown. Methods: Spatial and temporal patterns of gene expression were analyzed by quantitative reverse transcription (qRT)-polymerase chain reaction (PCR) and GUS histochemical assay. In planta transformation in heterologous Arabidopsis was generated by floral dipping using Agrobacterium tumefaciens (C58C1). Protein localization was analyzed by confocal microscopy via fluorescent protein tagging. Results: PgUGT72AL1 was highly expressed in the rhizome, upper root, and youngest leaf compared with the other organs. GUS staining of the promoter: GUS fusion revealed high expression in different organs, including axillary leaf branch. Overexpression of PgUGT72AL1 resulted in a fused organ in the axillary leaf branch. Conclusion: PgUGT72AL1, which is phylogenetically close to PgUGT71A27, is involved in the production of ginsenoside compound K. Considering that compound K is not reported in raw ginseng material, further characterization of this gene may shed light on the biological function of ginsenosides in ginseng plant growth and development. The organ fusion phenotype could be caused by the defective growth of cells in the boundary region, commonly regulated by phytohormones such as auxins or brassinosteroids, and requires further analysis.

• Mass Spectrometry Letters
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• v.9 no.2
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• pp.41-45
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• 2018

Ginseng (Panax ginseng Meyer) has been used as traditional herbal drug in Asian countries. Ginsenosides are major components having pharmacological and biological efficacy like anti-inflammatory, anti-diabetic and anti-tumor effects. To control the quality of the components in diverse ginseng products, we developed a new quantitative method using LC-MS/MS for 13 ginsenosides; Rb1, Rb2, Rc, Rd, Re, Rf, 20(S)-Rh1, 20(S)-Rh2, Rg1, 20(S)-Rg3, F1, F2, and compound K. This method was successfully validated for linearity, precision, and accuracy. This quantification method applied in four representative ginseng products; fresh ginseng powder, white ginseng powder, red ginseng extract powder, and red ginseng extract. Here the amounts of the 13 ginsenosides in the various type of ginseng samples could be analyzed simultaneously and expected to be suitable for quality control of ginseng products.