• Journal of Ginseng Research
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• v.44 no.1
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• pp.44-49
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• 2020

Background: Salting-out extraction (SOE) had been developed as a special branch of aqueous two-phase system recently. So far as we know, few reports involved in extracting ginsenosides with SOE because of the lower recovery caused by the unique solubility and surface activity of ginsenosides. A new SOE method for rapid pretreatment of ginsenosides from the enzymatic hydrolysates of Panax quinquefolium was established in this article. Methods: The SOE system comprising ethanol and sodium carbonate was selected to extract ginsenosides from the enzymatic hydrolysates of Panax quinquefolium, and HPLC was applied to analyze the ginsenosides. Results: The optimized extraction conditions were as follows: the aqueous two-phase extraction system comprising ethanol, sodium carbonate, ethanol concentration of 41.51%, and the mass percent of sodium carbonate of 7.9% in the extraction system under the experimental condition. Extraction time had minor influence on extraction efficiency of ginsenosides. The results also showed that the extraction efficiencies of three ginsenosides were all more than 90.0% only in a single step. Conclusion: The proposed method had been successfully applied to determine ginsenosides in enzymatic hydrolysate and demonstrated as a powerful technique for separating and purifying ginsenosides in complex samples.

• Journal of Ginseng Research
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• v.43 no.1
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• pp.105-115
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• 2019

Background: Ginsenosides with less sugar moieties may exhibit the better adsorptive capacity and more pharmacological activities. Methods: An efficient method for the separation of four minor saponins, including gypenoside XVII, notoginsenoside Fe, ginsenoside Rd2, and notoginsenoside Fd, from Panax notoginseng leaves (PNL) was established using biotransformation, macroporous resins, and subsequent preparative high-performance liquid chromatography. Results: The dried PNL powder was immersed in the distilled water at $50^{\circ}C$ for 30 min for converting the major saponins, ginsenosides Rb1, Rc, Rb2, and Rb3, to minor saponins, gypenoside XVII, notoginsenoside Fe, ginsenoside Rd2, and notoginsenoside Fd, respectively, by the enzymes present in PNL. The adsorption characteristics of these minor saponins on five types of macroporous resins, D-101, DA-201, DM-301, X-5, and S-8, were evaluated and compared. Among them, D-101 was selected due to the best adsorption and desorption properties. Under the optimized conditions, the fraction containing the four target saponins was separated by D-101 resin. Subsequently, the target minor saponins were individually separated and purified by preparative high-performance liquid chromatography with a reversed-phase column. Conclusion: Our study provides a simple and efficient method for the preparation of these four minor saponins from PNL, which will be potential for industrial applications.

• Journal of Microbiology and Biotechnology
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• v.29 no.2
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• pp.222-229
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• 2019

Korean ginseng (Panax ginseng Meyer) was processed by drying, steaming, or puffing, and the effects of these processes on the ginsenoside profile were investigated. The main root of 4-year-old raw Korean ginseng was dried to produce white ginseng. Steaming, followed by drying, was employed to produce red or black ginseng. In addition, these three varieties of processed ginseng were puffed using a rotational puffing gun. Puffed ginseng showed significantly higher extraction yields of ginsenosides (49.87-58.60 g solid extract/100 g of sample) and crude saponin content (59.40-63.87 mg saponin/g of dried ginseng) than non-puffed ginseng, respectively. Moreover, puffing effectively transformed the major ginsenosides (Rb1, Rb2, Rc, Rd, Re, and Rg1) of ginseng into minor ones (F2, Rg3, Rk1, and Rg5), comparable to the steaming process effect on the levels of the transformed ginsenosides. However, steaming takes much longer (4 to 36 days) than puffing (less than 30 min) for ginsenoside transformation. Consequently, puffing may be an effective and economical technique for enhancing the extraction yield and levels of minor ginsenosides responsible for the major biological activities of ginseng.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.477-479
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• 2018

A Gram-positive, rod-shaped, ivory colored, and motile, Lactobacillus koreensis 26-25 was isolated from Korean kimchi. Strain 26-25 showed the ability of conversion from major ginsenosides into minor ginsenosides for which whole genome was sequenced. The whole genome sequence of Lactobacillus koreensis 26-25 consisted of one circular chromosome comprised of 3,006,812 bp, with a DNA G + C content of 49.23%. The whole genome analysis of strain 26-25 showed many glycosides hydrolase genes, which may contribute to identify the genes responsible for transformation of major ginsenosides into minor ginsenosides for its high pharmacological effects.

• Journal of Ginseng Research
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• v.42 no.4
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• pp.504-511
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• 2018

Background: The biological activities of ginseng saponins (ginsenosides) are associated with type, number, and position of sugar moieties linked to aglycone skeletons. Deglycosylated minor ginsenosides are known to be more biologically active than major ginsenosides. Accordingly, the deglycosylation of major ginsenosides can provide the multibioactive effects of ginsenosides. The purpose of this study was to transform ginsenoside Rb2, one of the protopanaxadiol-type major ginsenosides, into minor ginsenosides using ${\beta}$-glycosidase (BG-1) purified from Armillaria mellea mycelium. Methods: Ginsenoside Rb2 was hydrolyzed by using BG-1; the hydrolytic properties of Rb2 by BG-1 were also characterized. In addition, the influence of reaction conditions such as reaction time, pH, and temperature, and transformation pathways of Rb2, Rd, F2, compound O (C-O), and C-Y by treatment with BG-1 were investigated. Results: BG-1 first hydrolyzes 3-O-outer ${\beta}$-$\text\tiny{D}$-glucoside of Rb2, then 3-O-${\beta}$-$\text\tiny{D}$-glucoside of C-O into C-Y. C-Y was gradually converted into C-K with a prolonged reaction time, but the pathway of Rb2 ${\rightarrow}$ Rd ${\rightarrow}$ F2 ${\rightarrow}$ C-K was not observed. The optimum reaction conditions for C-Y and C-K formation from Rb2 by BG-1 were pH 4.0-4.5, temperature $45-60^{\circ}C$, and reaction time 72-96 h. Conclusion: ${\beta}$-Glycosidase purified from A. mellea mycelium can be efficiently used to transform Rb2 into C-Y and C-K. To our best knowledge, this is the first result of transformation from Rb2 into C-Y and C-K by basidiomycete mushroom enzyme.

• 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.

• Journal of Ginseng Research
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• v.38 no.1
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• pp.47-51
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• 2014

The transformation of ginsenoside Rb1 into a specific minor ginsenoside using Aspergillus niger KCCM 11239, as well as the identification of the transformed products and the pathway via thin layer chromatography and high performance liquid chromatography were evaluated to develop a new biologically active material. The conversion of ginsenoside Rb1 generated Rd, Rg3, Rh2, and compound K although the reaction rates were low due to the low concentration. In enzymatic conversion, all of the ginsenoside Rb1 was converted to ginsenoside Rd and ginsenoside Rg3 after 24 h of incubation. The crude enzyme (b-glucosidase) from A. niger KCCM 11239 hydrolyzed the ${\beta}$-($1{\rightarrow}6$)-glucosidic linkage at the C-20 of ginsenoside Rb1 to generate ginsenoside Rd and ginsenoside Rg3. Our experimental demonstration showing that A. niger KCCM 11239 produces the ginsenoside-hydrolyzing b-glucosidase reflects the feasibility of developing a specific bioconversion process to obtain active minor ginsenosides.

• Journal of Ginseng Research
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• v.39 no.4
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• pp.392-397
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• 2015

Background: Red ginseng (RG) is processed from Panax ginseng via several methods including heat treatment, mild acid hydrolysis, and microbial conversion to transform the major ginsenosides into minor ginsenosides, which have greater pharmaceutical activities. During the fermentation process using microbial strains in a machine for making red ginseng, a change of composition occurs after heating. Therefore, we confirmed that fermentation had occurred using only microbial strains and evaluated the changes in the ginsenosides and their chemical composition. Methods: To confirm the fermentation by microbial strains, the fermented red ginseng was made with microbial strains (w-FRG) or without microbial strains (n-FRG), and the fermentation process was performed to tertiary fermentation. The changes in the ginsenoside composition of the self-manufactured FRG using the machine were evaluated using HPLC, and the 20 ginsenosides were analyzed. Additionally, we investigated changes of the reducing sugar and polyphenol contents during fermentation process. Results: In the fermentation process, ginsenosides Re, Rg1, and Rb1 decreased but ginsenosides Rh1, F2, Rg3, and Compound Y (C.Y) increased in primary FRG more than in the raw ginseng and RG. The content of phenolic compounds was high in FRG and the highest in the tertiary w-FRG. Moreover, the reducing sugar content was approximately three times higher in the tertiary w-FRG than in the other n-FRG. Conclusion: As the results indicate, we confirmed the changes in the ginsenoside content and the role of microbial strains in the fermentation process.

• Journal of Ginseng Research
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• v.41 no.3
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• pp.379-385
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• 2017

In this study, white ginseng was used as the raw material, which was fermented with Paecilomyces hepiali through solid culture medium, to produce ginsenosides with modified chemical composition. The characteristic chemical markers of the products thus produced were investigated using rapid resolution liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (RRLC-QTOF-MS). Chemical profiling data were obtained, which were then subjected to multivariate statistical analysis for the systematic comparison of active ingredients in white ginseng and fermented ginseng to understand the beneficial properties of ginsenoside metabolites. In addition, the effects of these components on biological activity were investigated to understand the improvements in the phagocytic function of macrophages in zebrafish. According to the established RRLC-QTOF-MS chemical profiling, the contents in ginsenosides of high molecular weight, especially malonylated protopanaxadiol ginsenosides, were slightly reduced due to the fermentation, which were hydrolyzed into rare and minor ginsenosides. Moreover, the facilitation of macrophage phagocytic function in zebrafish following treatment with different ginseng extracts confirmed that the fermented ginseng is superior to white ginseng. Our results prove that there is a profound change in chemical constituents of ginsenosides during the fermentation process, which has a significant effect on the biological activity of these compounds.

• Journal of Ginseng Research
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• v.35 no.3
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• pp.375-383
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• 2011

In the previous report, we have demonstrated that ginsenoside Rc, one of major ginsenosides, is a major component for the restoration for normal growth of worms in cholesterol-deprived medium. In the present study, we further investigated the roles of minor ginsenosides, such as ginsenoside $Rh_1$ and $Rh_2$, ginsenoside metabolites such as compound K (CK), protopanaxadiol (PPD), and protopanaxatriol (PPT) and ginsenoside epimers such as 20(R)- and 20(S)-ginsenoside $Rg_3$ in cholesterol-deprived medium. We found that ginsenoside $Rh_1$ almost restored normal growth of worms in cholesterol-deprived medium in F1 generation. However, supplement of ginsenoside $Rh_2$ caused a suppression of worm growths in cholesterol-deprived medium. In addition, CK and PPD also slightly restored normal growth of worms in cholesterol-deprived medium but PPT not. In experiments using ginsenoside epimers, supplement of 20(S)- but not 20(R)-ginsenoside $Rg_3$ in cholesterol-deprived medium also almost restored worm growth. These results indicate that the absence or presence of carbohydrate component at backbone of ginsenoside, the number of carbohydrate attached at carbon-3, and the position of hydroxyl group at carbon-20 of ginsenoside might plays important roles in restoration of worm growth in cholesterol-deprived medium.