• Journal of Ginseng Research
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• 제41권4호
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• pp.524-530
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• 2017

Background: Panax ginseng is a physiologically active plant widely used in traditional medicine that is characterized by the presence of ginsenosides. Rb1, a major ginsenoside, is used as the starting material for producing ginsenoside derivatives with enhanced pharmaceutical potentials through chemical, enzymatic, or microbial transformation. Methods: To investigate the bioconversion of ginsenoside Rb1, we prepared kimchi originated bacterial strains Leuconostoc mensenteroides WiKim19, Pediococcus pentosaceus WiKim20, Lactobacillus brevis WiKim47, Leuconostoc lactis WiKim48, and Lactobacillus sakei WiKim49 and analyzed bioconversion products using LC-MS/MS mass spectrometer. Results: L. mesenteroides WiKim19 and Pediococcus pentosaceus WiKim20 converted ginsenoside Rb1 into the ginsenoside Rg3 approximately five times more than Lactobacillus brevis WiKim47, Leuconostoc lactis WiKim48, and Lactobacillus sakei WiKim49. L mesenteroides WIKim19 showed positive correlation with b-glucosidase activity and higher transformation ability of ginsenoside Rb1 into Rg3 than the other strains whereas, P. pentosaceus WiKim20 showed an elevated production of Rb3 even with lack of b-glucosidase activity but have the highest acidity among the five lactic acid bacteria (LAB). Conclusion: Ginsenoside Rg5 concentration of five LABs have ranged from ${\sim}2.6{\mu}g/mL$ to $6.5{\mu}g/mL$ and increased in accordance with the incubation periods. Our results indicate that the enzymatic activity along with acidic condition contribute to the production of minor ginsenoside from lactic acid bacteria.

• Journal of Ginseng Research
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• 제34권4호
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• pp.288-295
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• 2010

Ginsenosides are the principal components responsible for the pharmacological and biological activities of ginseng. Ginsenoside Rd was transformed into compound K using cell-free extracts of food microorganisms, with Lactobacillus pentosus DC101 isolated from kimchi (traditional Korean fermented food) used for this conversion. The optimum time for the conversion was about 72 h at a constant pH of 7.0 and an optimum temperature of about $30^{\circ}C$. The transformation products were identified by thin-layer chromatography and high-performance liquid chromatography, and their structures were assigned using nuclear magnetic resonance analysis. Generally, ginsenoside Rd was converted into ginsenoside F2 by 36 h post-reaction. Consequently, over 97% of ginsenoside Rd was decomposed and converted into compound K by 72 h post-reaction. The bioconversion pathway to produce compound K is as follows: ginsenoside Rd$\rightarrow$ginsenoside F2$\rightarrow$compound K.

• Mycobiology
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• 제42권3호
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• pp.256-261
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• 2014

A ${\beta}$-glucosidase producing yeast strain was isolated from Korean traditional rice wine. Based on the sequence of the YCL008c gene and analysis of the fatty acid composition, the isolate was identified as Saccharomyces cerevisiae strain HJ-014. S. cerevisiae HJ-014 produced ginsenoside Rd, $F_2$, and compound K from the ethanol extract of red ginseng. The production was increased by shaking culture, where the bioconversion efficiency was increased 2-fold compared to standing culture. The production of ginsenoside $F_2$ and compound K was time-dependent and thought to proceed by the transformation pathway of: red ginseng extract ${\rightarrow}Rd{\rightarrow}F_2{\rightarrow}$ compound K. The optimum incubation time and concentration of red ginseng extract for the production of compound K was 96 hr and 4.5% (w/v), respectively.

• Journal of Microbiology and Biotechnology
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• 제32권4호
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• pp.447-457
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• 2022

Notoginsenoside R1 and ginsenoside Rg1 are the main active ingredients of Panax notoginseng, exhibiting anti-fatigue, anti-tumor, anti-inflammatory, and other activities. In a previous study, a GH39 β-xylosidase Xln-DT was responsible for the bioconversion of saponin, a natural active substance with a xylose group, with high selectivity for cleaving the outer xylose moiety of notoginsenoside R1 at the C-6 position, producing ginsenoside Rg1 with potent anti-fatigue activity. The optimal bioconversion temperature, pH, and enzyme dosage were obtained by optimizing the transformation conditions. Under optimal conditions (pH 6.0, 75℃, enzyme dosage 1.0 U/ml), 1.0 g/l of notoginsenoside R1 was converted into 0.86 g/l of ginsenoside Rg1 within 30 min, with a molar conversion rate of approximately 100%. Furthermore, the in vivo anti-fatigue activity of notoginsenoside R1 and ginsenoside Rg1 were compared using a suitable rat model. Compared with the control group, the forced swimming time to exhaustion was prolonged in mice by 17.3% in the Rg1 high group (20 mg/kg·d). Additionally, the levels of hepatic glycogen (69.9-83.3% increase) and muscle glycogen (36.9-93.6% increase) were increased. In the Rg1 group, hemoglobin levels were also distinctly increased by treatment concentrations. Our findings indicate that treatment with ginsenoside Rg1 enhances the anti-fatigue effects. In this study, we reveal a GH39 β-xylosidase displaying excellent hydrolytic activity to produce ginsenoside Rg1 in the pharmaceutical and food industries.

• 한국미생물·생명공학회지
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• 제42권4호
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• pp.347-353
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• 2014

본 연구는 인삼의 주요성분인 ginsenoside Rb1으로부터 보다 높은 생리기능성을 갖는 것으로 알려져 있는 compound K를 생산하기 위하여 Aspergillus usamii KCTC 6954에서 유래된 ${\beta}$-glucosidase를 사용하여 생물전환을 실시하였다. 15일 동안의 배양 중, 효소활 성 측정은 ${\rho}$-nitrophenyl-${\beta}$-glucopyranoside를 기질로 하여 분해 생성되는 ${\rho}$-nitrophenol (${\rho}NP$)을 비색계로 측정함으로써 실시되었다. 그 결과로서, 균주의 성장 속도는 접종 후 6일 후 최대로 나타났으며 이때의 ${\beta}$-glucosidas 활성도는 $175.93{\mu}M\;ml^{-1}min^{-1}$로 나타났다. 또 한 효소 반응의 최적 조건은 pH 6.0 이내에서는 $60^{\circ}C$인 것으로 나타났다. 배양 중 ginsenosides 분석 결과, 배양 9일 후에는 Rb1는 Rd 로 전환되고 15 days 후에는 compound K로 순차적으로 전환되는 것으로 나타났다. 효소반응에 있어서는 Rb1는 1시간 이내에 ginsenoside Rd로 전환되었고 8시간 이후에 최종산물인 compound K가 측정되었다. 본 연구결과로부터 Rb1으로부터 주요 생물학적 전환 경로는 $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}$compound K로 나타났으며 이는 차후 Rd나 compound K와 같이 강한 생리기능성을 갖지만 자연에 미 량 존재하는 물질의 대량생산에 응용될 수 있을 것으로 기대된다.

• 한국자원식물학회지
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• 제24권6호
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• pp.712-718
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• 2011

Ginseng (Panax ginseng) is frequently used in Asian countries as a traditional medicine. The major components of ginseng are ginsenosides. Among these, ginsenoside compound K has been reported to prevent the formation of malignancy and metastasis of cancer by blocking the formation of tumor and suppressing the invasion of cancer cells. In this study, ginsenoside $Rb_1$ was converted into compound K, via secreted ${\beta}$-glucosidase enzyme from the Leuconostoc lactis DC201 isolated, which was extracted from Kimchi. The strain DC201 was suspended and cultured in MRS broth at $37^{\circ}C$. Subsequently, the residue from the cultured broth supernatant was precipitated with EtOH and then dissolved in 20 mM sodium phosphate buffer (pH 6.0) to obtain an enzyme liquid. Meanwhile, the crude enzyme solution was mixed with ginsenoside $Rb_1$ at a ratio of 1:4 (v/v).The reaction was carried out at $30^{\circ}C$ and 190 rpm for 72 hours, and then analyzed by TLC and HPLC. The result showed that ginsenoside Rb1 was transformed into compound K after 72 hours post reaction.

• Journal of Ginseng Research
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• 제38권1호
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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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• 제40권2호
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• pp.121-126
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• 2016

Background: Ginsenoside F1, a pharmaceutical component of ginseng, is known to have antiaging, antioxidant, anticancer, and keratinocyte protective effects. However, the usage of ginsenoside F1 is restricted owing to the small amount found in Korean ginseng. Methods: To enhance the production of ginsenoside F1 as a 10 g unit with high specificity, yield, and purity, an enzymatic bioconversion method was developed to adopt the commercial enzyme Cellulase KN from Aspergillus niger with food grade, which has ginsenoside-transforming ability. The proposed optimum reaction conditions of Cellulase KN were pH 5.0 and $50^{\circ}C$. Results: Cellulase KN could effectively transform the ginsenosides Re and Rg1 into F1. A scaled-up biotransformation reaction was performed in a 10 L jar fermenter at pH 5.0 and $50^{\circ}C$ for 48 h with protopanaxatriol-type ginsenoside mixture (at a concentration of 10 mg/mL) from ginseng roots. Finally, 13.0 g of F1 was produced from 50 g of protopanaxatriol-type ginsenoside mixture with $91.5{\pm}1.1%$ chromatographic purity. Conclusion: The results suggest that this enzymatic method could be exploited usefully for the preparation of ginsenoside F1 to be used in cosmetic, functional food, and pharmaceutical industries.

• Journal of Ginseng Research
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• 제43권1호
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• pp.20-26
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• 2019

Background: Ginsenosides, which are bioactive components in ginseng, can be converted to smaller compounds for improvement of their pharmacological activities. The conversion methods include heating; acid, alkali, and enzymatic treatment; and microbial conversion. The aim of this study was to determine the bioconversion of ginsenosides in fermented red ginseng extract (FRGE). Methods: Red ginseng extract (RGE) was fermented using Lactobacillus plantarum KCCM 11613P. This study investigated the ginsenosides and their antioxidant capacity in FRGE using diverse methods. Results: Properties of RGE were changed upon fermentation. Fermentation reduced the pH value, but increased the titratable acidity and viable cell counts of lactic acid bacteria. L. plantarum KCCM 11613P converted ginsenosides $Rb_2$ and $Rb_3$ to ginsenoside Rd in RGE. Fermentation also enhanced the antioxidant effects of RGE. FRGE reduced 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and reducing power; however, it improved the inhibition of ${\beta}$-carotene and linoleic acid oxidation and the lipid peroxidation. This suggested that the fermentation of RGE is effective for producing ginsenoside Rd as precursor of ginsenoside compound K and inhibition of lipid oxidation. Conclusion: This study showed that RGE fermented by L. plantarum KCCM 11613P may contribute to the development of functional food materials.

• Journal of Microbiology and Biotechnology
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• 제17권12호
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• pp.1937-1943
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• 2007

A new strain, GS603, having ${\beta}$-glucosidase activity was isolated from soil of a ginseng field, and its ability to convert major ginsenoside $Rb_1$ to minor ginsenoside or gypenoside was studied. Strain GS603 was identified as an Intrasporangium species by phylogenetic analysis and showed high ginsenoside-converting activity in LB and TSA broth but not in nutrient broth. The culture broth of the strain GS603 could convert ginsenoside $Rb_1$i into two metabolites, which were analyzed by TLC and HPLC and shown to be the minor ginsenoside $F_2$ and gypenoside XVII by NMR.