• 한국식품과학회지
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• 제29권6호
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• pp.1327-1329
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• 1997

인삼으로부터 사포닌 화합물의 신속한 추출방법을 모색하기 위하여 waring blonder 와 유기용매를 이용한 새로운 추출방법을 개발하였다. 본 방법은 기존의 인삼 증류추출방법과 비교해볼 때 6개의 주종 사포닌($ginsenoside-Rb_2$, $ginsenoside-Rb_1$, ginsenoside-Rc, ginsenoside-Rd, ginsenoside-Re, $ginsenoside-Rg_1$) 함량이 유사하여 유의성이 있는 것으로 생각된다. 유기용매는 메탄올과 클로로포름을 7:3의 비율로 사용하였을 때 사포닌 화합물이 잘 추출되었다. 5개의 주종사포닌을 합한 전체 사포닌 함량은 본 방법에서는 2.41% 이었고 기존의 방법 에서는 2.54%이었다. 그러나 조사포닌의 함량은 본 방법이 기존의 방법보다 높은 것으로 나타났다.

• Journal of Ginseng Research
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• 제40권4호
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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.

• 한국식품영양학회지
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• 제23권3호
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• pp.405-410
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• 2010

인삼의 저장 가공 방법을 개발하기 위해 마이크로파 단독처리로 수삼을 처리한 후 일반 성분, 색도, 밀도, 사포닌의 조성 및 미세구조 등 품질 특성을 조사하였다. 마이크로웨이브 출력을 100 Watt와 200 Watt에서 각각 1시간과 3시간 동안 수삼을 처리하고 $60^{\circ}C$ 열풍건조기에서 96시간 건조한 후 성분 함량을 측정하였다. 수삼의 초기 수분 함량인 76.26%에서 마이크로파 가열 처리 조건에 따라 13.12~10.77%까지 건조되었다. 마이크로파로 처리한 수삼의 회분 함량은 처리 과정에 따른 차이는 보이지 않았다. 지방의 경우 마이크로파 출력과는 상관없이 처리 시간에 따라 조금씩 감소하였다. 단백질 함량은 마이크로파 가열 후 건조하기 전 수삼의 단백질 함량과 비교하여 큰 차이를 나타내지 않았다. 마이크로파로 처리한 수삼의 총 식이섬유 함량은 대조군보다 증가함을 보이고 있으나, 처리 시간이나 출력과의 유의성은 보이지 않는다. 처리군의 색도는 대조군에 비해 더 어두운 색을 나타내었다. 마이크로파 가열 처리 후 인삼의 색도의 변화는 출력보다 가열 처리 시간에 따라 변하는 양상을 보였다. 마이크로파 가열 처리 후 수삼의 밀도는 감소하는 경향을 보였고, 가열 시간이나 출력에 따른 뚜렷한 차이는 찾을 수 없었다. 사포닌 ginsenoside-$Rb_1$과 $Rb_2+Rb_3$, Rc, Rd, Re, Rf, $Rg_1$, $Rg_2+Rh_1$, $Rg_3$의 함량은 대조군에 비해 감소하는 경향을 보였고, 100 Watt에서 가열한 처리구들보다 200 Watt에서 가열한 처리구들의 함량이 높은 값을 나타냈다. 예외적으로 200 Watt에서 가열했을 때, ginsenoside-$Rb_2+Rb_3$와 ginsenoside-Rc의 함량은 대조군에 비해 증가하였다. 미세구조는 마이크로파 건조 후 더 치밀한 조직이 관찰되었다.

• Journal of Microbiology and Biotechnology
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• 제18권6호
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• pp.1081-1089
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• 2008

A novel ginsenoside-hydrolyzing ${\beta}$-glucosidase was purified from Paecilomyces Bainier sp. 229 by a combination of Q-Sepharose FF, phenyl-Sepharose CL-4B, and CHT ceramic hydroxyapatite column chromatography. The purified enzyme was a monomeric protein with a molecular mass estimated to be 115 kDa. The optimal enzyme activity was observed at pH 3.5 and $60^{\circ}C$. It was highly stable within pH 3-9 and at temperatures lower than $55^{\circ}C$. The enzyme was specific to ${\beta}$-glucoside. The order of enzyme activities against different types of ${\beta}$-glucosidic linkages was ${\beta}$-(1-6)>${\beta}$-(1-2)>${\beta}$-(1-4). The enzyme converted ginsenoside Rb1 to CK specifically and efficiently. An 84.3% amount of ginsenoside Rb1, with an initial concentration of 2 mM, was converted into CK in 24 h by the enzyme at $45^{\circ}C$ and pH 3.5. The hydrolysis pathway of ginsenoside Rb1 by the enzyme was $Rb1{\to}Rd{\to}F2{\to}CK$. Five tryptic peptide fragments of the enzyme were identified by a newly developed de novo sequencing method of post-source decay (PSD) matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. By comparing the five identified peptide sequences with the NCBI database, this purified ${\beta}$-glucosidase proves to be a new protein that has not been reported before.

• 한국식품영양과학회지
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• 제41권12호
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• pp.1753-1757
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• 2012

LC-MS/MS를 사용함으로써 인체 혈장에서 ginsenoside Rb1의 분석법을 개발하고 검증하였다. 유속 0.9 mL/min에 이동상 0.1% formic acid가 첨가된 water와 methanol을 사용하여 기울기 용리 조건으로 설정하였으며 사용한 분석 칼럼은 C 18($4.6mm{\times}150mm$, particle size 5 ${\mu}m$)을 사용하여 분리하였다. MRM(multiple reaction monitoring) 방법의 전기 분무 이온화 이온 분석기로 모니터링 하여 분석하였다. 인체 혈장 샘플은 액체-액체 추출방법에 의해 acetone과 water가 섞인 용액으로 추출하였다. 이 분석의 검량선 범위는 10~500 ng/mL이며 상관계수는 0.9995를 나타냈다. 일내, 일간의 정밀성 농도범위는 상관계수 5.8% 그리고 정확성은 96.0~104.6%로 나타났다. 이 LC-MS/MS를 이용한 인체 혈장의 ginsenoside Rb1의 연구가 약동학 연구에 적용할 수 있을 거라 생각한다.

• 생명과학회지
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• 제29권5호
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• pp.514-523
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• 2019

인삼(Panax ginseng C. A. Meyer)의 사포닌 진세노사이드 Rb1이 처리된 인간 피부각질세포 HaCaT에서 microarray 분석 및 발현이 증가된 세포사멸 반응에 대한 작용기전을 연구하였다. HaCaT 세포에 진세노사이드 Rb1의 처리로 세포사멸, 유사분열 세포주기의 G2/M 전이, 세포분열, 핵분열, 그리고 단백질 수송 등의 작용기전에 관여하는 유전자들이 2 배 이상 발현이 증가된 것으로 나타났으며, DNA 수선, 감수 핵분열, 그리고 세포외기질 체계 등의 작용기전에 관여하는 유전자들은 2 배 이상 발현이 감소된 것으로 나타났다. 특히 세포사멸 신호전달은 FAS와 PLA2G4A를 경유하는 것으로 나타났으며, 이들 유전자의 상위 조절자로 STAT3가 예측되었다. 세포사멸 반응 경유 유전자 FAS와 PLA2G4A의 활성을 qPCR로 확인한 결과, FAS 유전자는 $10{\mu}g/ml$의 진세노사이드 Rb1를 24시간 동안 처리하였을 경우 약 2 배의 발현 증가와, PLA2G4A 유전자는 6시간 처리부터 약 2 배로 증가되어 24시간 동안 처리시 2 배 이상의 유전자 발현이 증가되었다. 한편 STAT3-siRNA를 이용한 knock-down 실험에서 FAS의 발현 감소와 PLA2G4A의 발현 증가로 상위 조절자 STAT3로부터 FAS 만을 경유하는 것을 알 수 있었다. 이상의 결과 진세노사이드 Rb1의 처리에 의해 상위 조절자인 STAT3는 FAS를 경유하여 세포사멸을 유도하는 것임을 알 수 있다.

• 동의생리병리학회지
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• 제22권6호
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• pp.1557-1561
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• 2008

Red ginseng possibly has new ingredients converted during steaming and dry process from fresh ginseng. Kujeungkupo method which means 9 repetitive steamings and dryings process was used for the production of red ginseng from 6-year old ginseng roots. Saponin was extracted from each red ginseng produced at the 1st, 3rd, 5th, 7th, and 9th during the steaming and drying treatment, and we analyzed saponin content with TLC. Minor saponins, such as ginsenoside-Rg3, -Rh2, compound K, and F2, increased as the process time of steaming and drying, but major saponins (ginsenoside-Rb1, -Rb2, -Rc, -Rd, -Re, -Rf, -Rg1) were decreased. Major saponins were yet observed almost at the 1st process, then degraded as the increasing time of steaming and drying process. Especially, ginsenoside-Re and -Rg were observed as considerable amount after the 1st treatment, but there were no trace of them after the 9th treatment. Ginsenoside-Rg1, -Rb2, and -Rb1 were also reduced remarkedly by 96.6%, 96%, and 92.3%, respectively. Minor saponins were increased significantly, especially for ginsenoside-Rg3 and ginsenoside-F2. These results suggest that Kujeungkupo method is the very useful method for the production of minor ginsenoside-Rg3 and -Rh2.

• Journal of Ginseng Research
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• 제45권2호
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• pp.287-294
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• 2021

Background: Ginsenoside Rb1 (G-Rb1), one of the major active compounds in Panax ginseng, has already been shown to reduce inflammation in various diseases. Osteoarthritis (OA) has traditionally been considered a degenerative disease with degradation of joint articular cartilage. However, recent studies have shown the association of inflammation with OA. In the present study, we investigated whether Rb1 had an antiinflammatory effect on monoiodoacetate (MIA)-induced OA in ovariectomized rats as a model of postmenopausal arthritis. Methods: G-Rb1 at a dosage of 3 and 10 ㎍/kg body weight was administered every 3 days intraarticularly for a period of 4 weeks to observe antiarthritic effects. Diclofenac (10 mg/kg) served as a positive control. Results: The administration of Rb1 significantly ameliorated OA inflammatory symptoms and reduced serum levels of inflammatory cytokines. Furthermore, G-Rb1 administration considerably enhanced the expression of bone morphogenetic protein-2 and collagen 2A and reduced the levels of matrix metalloproteinase-13 genes, indicating a chondroprotective effect of G-Rb1. G-Rb1 also significantly reduced the expression of several inflammatory cytokines/chemokines (interferon gamma (IFN-γ), monocyte chemoattractant protein-1 (MCP-1)/CCL-2, interleukin [IL]-1β, and IL-6). Histological analysis demonstrated that G-Rb1 significantly attenuated the pathological changes in MIA-induced OA in ovariectomized rats. Safranin O and toluidine blue staining further demonstrated that G-Rb1 effectively prevented the degradation of cartilage and glycosaminoglycans, respectively. Conclusion: Overall, our results suggest that G-Rb1 exerts cartilage protective effect on MIA-induced ovariectomized OA rats, by inhibiting inflammatory mediators such as IL-6, IL-1β, MCP-1/CCL-2, cyclooxygenase-2 (COX-2), and prostaglandin E₂ (PGE2). These results shed a light on possible therapeutic application of G-Rb1 in OA.

• BMB Reports
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• 제32권4호
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• pp.339-344
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• 1999

We studied the effects of ginsenosides in immature rats based upon the previous results that ginseng has a suppressive or anticonvulsive activity. To examine the suppressive effect of ginsenosides on kainic acid-induced seizures, the severities and frequencies were observed for 4 h after injection of kainic acid (KA; i.p., 2 mg/kg b.w.) using 10-day-old male Sprague-Dawley rats ($22{\pm}2\;g$). Protopanaxadiol saponins such as ginsenoside-Rb1 (Rb1), ginsenoside-Rb2 (Rb2), ginsenoside-Rc (Rc), and ginsenoside-Rd(Rd) generally reduced the seizure activities while protopanaxatriol saponins such as ginsenoside-Rg1 (Rg1) and ginsenoside-Re (Re) rather increased stereotypic "paddling-like" movements. When vinyl-GABA (v-G) was injected together with Rb1 or Rc, KA-induced seizure severities were additionally reduced only by the injection of Rc, but not by Rb1. The level of gamma isozyme of protein kinase C (PKC-${\gamma}$) in the hippocampus increased about three times as much as that of normal rats at 4 h after KA injection. The increased level of PCK-${\gamma}$ by KA was significantly reduced to about 35% by the coinjection with v-G alone, but it was not changed by v-G together with Rb1 or Rc. The increased level of PKC-${\gamma}$ at 4 h after injection of KA was not consistent with the reduction of seizure severities between Rb1 and Rc. These results suggest that Rc and Rb1 may reduce seizure severity independent of PKC-${\gamma}$ levels, and Rc may additionally act with v-G regarding the GABA metabolism during the stage of KA-induced seizures in the immature rats.

• Journal of Ginseng Research
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• 제45권2호
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• pp.334-343
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• 2021

Background: Gut microbiota mainly function in the biotransformation of primary ginsenosides into bioactive metabolites. Herein, we investigated the effects of three prebiotic fibers by targeting gut microbiota on the metabolism of ginsenoside Rb1 in vivo. Methods: Sprague Dawley rats were administered with ginsenoside Rb1 after a two-week prebiotic intervention of fructooligosaccharide, galactooligosaccharide, and fibersol-2, respectively. Pharmacokinetic analysis of ginsenoside Rb1 and its metabolites was performed, whilst the microbial composition and metabolic function of gut microbiota were examined by 16S rRNA gene amplicon and metagenomic shotgun sequencing. Results: The results showed that peak plasma concentration and area under concentration time curve of ginsenoside Rb1 and its intermediate metabolites, ginsenoside Rd, F2, and compound K (CK), in the prebiotic intervention groups were increased at various degrees compared with those in the control group. Gut microbiota dramatically responded to the prebiotic treatment at both taxonomical and functional levels. The abundance of Prevotella, which possesses potential function to hydrolyze ginsenoside Rb1 into CK, was significantly elevated in the three prebiotic groups (P < 0.05). The gut metagenomic analysis also revealed the functional gene enrichment for terpenoid/polyketide metabolism, glycolysis, gluconeogenesis, propanoate metabolism, etc. Conclusion: These findings imply that prebiotics may selectively promote the proliferation of certain bacterial stains with glycoside hydrolysis capacity, thereby, subsequently improving the biotransformation and bioavailability of primary ginsenosides in vivo.