• Journal of Ginseng Research
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• 제22권2호
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• pp.126-132
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• 1998

In the present study, we examined effects of ginseng saponins (ginsenosides) on pp60c-src protein tyrosine kinase (PTK) activity, intracellular calcium concentration ([$Ca^{2+}$]i), and cell proliferation in NIH3T3 cells. Eight different ginsenosides [ginsenoside-Rb1 (G-$Rb_1$), -$Rb_2$, -Rc, -Rd, -Re, -Rf, -$Rg_1$, -$Rg_2$) and ginseng total saponin (GTS) were used for these experiments. All ginsenosides and GTS tested stimulated the activation of $pp60^{c-src}$ kinase, and especially G-$Rb_1$,-Rd,-$Rg_1$, and -$Rg_1$ showed a higher stimulatory effect than others at 16.7 $\mu\textrm{g}$/ml of ginsenosides with a 18 hr-incubation, increasing the activity by 4.5, 3.5, 3.5, and 3.0-fold, respectively, over that of untreated control. In addition, both G-Rd and -$Rg_2$)Rg2 increased ($Ca^{2+}$), to 202 and 334 nM, respectively, about 2-3-fold above the basal level within 7min at 250 $\mu\textrm{g}$/yml of ginsenosides. The increases of ($Ca^{2+}$), were eliminated by Pretreatment of EGTA, an extracellular calcium chelator, suggtasting that they result from an influx of calcium ion from extracellular medium rather than an efflux from intracellular calcium store, endoplasmic reticulum (ER). All ginsenosides studied enhanced cell proliferation to 1.2-1.4-fold over that of untreated control at 5~250 $\mu\textrm{g}$/ml of concentrations. Interestingly the promotion of cell proliferation by ginsenosides corresponded with the activation of c-src kinase, which is an early step in the mitogenic signaling cascade. Taken together, we suggest that some ginsenosides may lead to cellProliferation via the activation of cellular signal transduction Pathway involving $pp60^{c-src}$ kinase.

• Journal of Ginseng Research
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• 제36권2호
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• pp.146-152
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• 2012

Panax ginseng (PG) is a globally utilized medicinal herb. The medicinal effects of PG are primarily attributable to ginsenosides located in the root and leaf. The leaves of PG are known to be rich in various bioactive ginsenosides, and the therapeutic effects of ginseng extract and ginsenosides have been associated with immunomodulatory and anti-inflammatory activities. We examined the effect of PG leaf extract and the isolated ginsenosides, on nuclear factor (NF)-${\kappa}B$transcriptional activity and target gene expression by applying a luciferase assay and reverse transcription polymerase chain reaction in tumor necrosis factor (TNF)-${\alpha}$-treated hepatocarcinoma HepG2 cells. Air-dried PG leaf extract inhibited TNF-${\alpha}$-induced NF-${\kappa}B$transcription activity and NF-${\kappa}B$-dependent cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) gene expression more efficiently than the steamed extract. Of the 10 ginsenosides isolated from PG leaves, Rd and Km most significantly inhibited activity in a dose-dependent manner, with $IC_{50}$ values of $12.05{\pm}0.82$ and $8.84{\pm}0.99\;{\mu}M$, respectively. Furthermore, the ginsenosides Rd and Km inhibited the TNF-${\alpha}$-induced expression levels of the COX-2 and iNOS gene in HepG2 cells. Air-dried leaf extracts and their chemical components, ginsenoside Rd and Km, are involved in the suppression of TNF-${\alpha}$-induced NF-${\kappa}B$ activation and NF-${\kappa}B$-dependent iNOS and COX-2 gene expression. Consequently, air-dried leaf extract from PG, and the purified ginsenosides, have therapeutic potential as anti-inflammatory.

• Journal of Applied Biological Chemistry
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• 제64권3호
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• pp.253-261
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• 2021

본 연구에서는 식물공장을 통해 생산된 새싹인삼을 크기에 따라 분류하고 이를 지상부와 지하부로 나누어 진세노사이드 함량과 항산화 활성을 비교하였다. 지상부의 경우 총 phenolic 함량은 중간 크기인 M에서 5.16 GAE mg/g로 가장 높았으며 가장 큰 크기인 L에서 2.23 GAE mg/g으로 가장 낮은 함량을 보였다. 지하부 역시 M 크기에서 가장 높은 함량을 보였으나, 큰 차이를 보이지 않았다. 한편, 총 flavonoid 함량 역시 지상부(5.16 RE mg/g) 와 지하부(1.28 RE mg/g) 모두 M 크기에서 높은 함량을 보였다. 지상부의 주요 진세노사이드는 Re (20.33-24.15 mg/g) > Rd (11.36-27.42 mg/g) > Rg1 (4.48-5.54 mg/g) 순 있었고 지하부는 Rb1 (5.09-8.61 mg/g) > Re (4.48-5.54 mg/g) > Rc (3.11-4.11 mg/g) 순 있었다. M 크기의 경우 Re와 Rd는 각각 지상부에서 24.15 mg/g과 27.42 mg/g 및 지하부에서 5.20 mg/g과 1.43 mg으로 약 4배와 19배 높은 함량을 보였다. 지상부에는 F3 및 Rh1이 검출되었으나, 지하부에서는 검출되지 않았다. DPPH (74.95%)와 ABTS (94.47%), hydroxyl (70.39%) 라디칼 소거 활성 및 FRAP (2.169) 활성은 다른 크기들보다 M 크기에서 가장 높은 활성을 보였다.

• Journal of Microbiology and Biotechnology
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• 제17권6호
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• pp.905-912
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• 2007

A novel ${\beta}-glucosidase$ gene, bglA, was isolated from uncultured soil bacteria and characterized. Using genomic libraries constructed from soil DNA, a gene encoding a protein that hydrolyzes a fluorogenic analog of cellulose, 4-methylumbelliferyl ${\beta}-D-cellobioside$ (MUC), was isolated using a microtiter plate assay. The gene, bglA, was sequenced using a shotgun approach, and expressed in E. coli. The deduced 55-kDa amino acid sequence for bglA showed a 56% identity with the family 1 glycosyl hydrolase Chloroflexus aurantiacus. BglA included two conserved family 1 glycosyl hydrolase regions. When using $p-nitrophenyl-{\beta}-D-glucoside$ (pNPG) as the substrate, the maximum activity of the purified ${\beta}-glucosidase$ exhibited at pH 6.5 and $55^{\circ}C$, and was enhanced in the presence of $Mn^{2+}$. The $K_m\;and\;V_{max}$ values for the purified enzyme with pNPG were 0.16 mM and $19.10{\mu}mol/min$, respectively. The purified BglA enzyme hydrolyzed both pNPG and $p-nitrophenyl-{\beta}-D-fucoside$. The enzyme also exhibited substantial glycosyl hydrolase activities with natural glycosyl substrates, such as sophorose, cellobiose, cellotriose, cellotetraose, and cellopentaose, yet low hydrolytic activities with gentiobiose, salicin, and arbutin. Moreover, BglA was able to convert the major ginsenoside $Rb_1$ into the pharmaceutically active minor ginsenoside Rd within 24 h.

• 한국약용작물학회지
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• 제22권1호
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• pp.17-22
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• 2014

Ginsenosides of roots in Panax ginseng were analyzed by metabolic-targeting HPLC using the partial least squares discriminant analysis (PLS-DA) and compared depending on sowing methods between direct seeding and transplanting method. Score plots derived from PLS-DA could identify the sowing method between the direct seeding and transplanting method in P. ginseng roots. The ginsenoside compounds were assigned as Rg1, Re, Rf, Rg2, Rb1, Rc, Rb2, Rb3, and Rd. Contents of Re, Rf, Rg2, Rb1, Rc, Rb3, and Rd of main roots produced from the transplanting method were relatively higher than those of samples produced from direct seeding method. Also, contents of Rg1, Re, Rf, Rg2, Rb1, Rc, Rb2, Rb3, and Rd of lateral roots from the transplanted samples were relatively higher than those of samples produced from direct seeding method. Therefore, HPLC with PLS-DA analysis can be a straightforward tool for identification of ginsenosides in main or lateral roots of P. ginseng obtained from two different seeding methods between direct and transplanting methods.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2020년도 춘계학술대회
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• pp.93-93
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• 2020

본 연구는 7년, 13년근 산양삼의 생육특성과 진세노사이드(G) 함량 간의 상관관계를 구명하기 위하여 수행되었다. 6개소의 산양삼의 생육특성을 조사한 결과, 뇌두길이, 뿌리길이, 생중량, 단면적, 표면적, 부피에 있어 13년근 산양삼이 7년근 산양삼에 비하여 유의적으로 높은 것을 확인하였다. 진세노사이드11종에 대한 함량은 G-Rb1, Rb2, Rc, Rd, Re, Rf, Rg1, Rg2 함량이 13년근 산양삼이 7년근 산양삼 보다 유의적으로 높은 수치를 확인하였다. 또한 산양삼과 인삼(재배삼) 진세노사이드 함량을 비교한 결과, 13년 산양삼에서 G-Rb1, Rd, Re, Rf, Rg1이 4년, 5년근 인삼(재배삼)에 비해 유의적으로 함량이 높은 것으로 확인되었다. 산양삼 연근별 생육특성과 진세노사이드 함량 간의 상관관계를 분석한 결과, G-Rb1, Rb2, Rc, Rd, Re, Rf, Rg1, Rg2 함량은 뇌두길이, 생중량, 단면적, 표면적, 부피와 유의정인 정의 상관관계를 보였으며, G-Rb1, Re, Rf, Rg2는 줄기직경과 부의 상관관계를 확인하였다. 본 연구는 산양삼의 7년근과 13년근을 대상으로 생육특성과 진세노사이드 함량 상관관계를 구명함으로써 연근에 따른 품질규격 정립에 유용한 정보를 제공 할 것으로 판단된다.

• 한국약용작물학회지
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• 제26권2호
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• pp.148-156
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• 2018

Background: Hot steaming is known to be effective in improving the biological activities of plant extracts by breaking down useful compounds to low molecular weight ones. Methods and Results: This study aimed to develop an optimal extraction and steam processing method for enhancing the low molecular ginsenoside contents of the adventitious roots culture of wild mountain ginseng. The total ginsenoside was optimally extracted when 70% EtOH was used at $50^{\circ}C$, whereas low molecule ginsenoside such as Rg2, Rh1, Rh4 and Rk1 could be extracted using 70% EtOH at $70^{\circ}C$. The adventitious roots culture of wild mountain ginseng is known to contain four major ginsenosides, i.e., Rb2, Rb1, Rg1 and Rd, however new ginsenosides Rg6, Rh4, Rg3, Rk1 and Rg5 were new abundantly obtaind after steam processing method was applied. The contents of total ginsenosides were the highest when thermal steam processing was conducted at $120^{\circ}C$ for 120 min. Unlike ginsenosides such as Rg1, Re, Rb1, Rc, Rb2, and Rh1, which decreased after steam processing, Rg3, Rk1, and Rg5 increased after thermal processing. Steam processing significanltly reduced the content of Rb1, increased that of Rg6 by about ten times than that in the adventitious roots culture of wild mountain ginseng. Conclusions: Our study showed that the optimal extraction and steam processing method increased the content of total ginsenosides and allowed the extraction of minor ginsenosides from major ones.

• Journal of Ginseng Research
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• 제40권1호
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• pp.47-54
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• 2016

Background: The roots of Panax ginseng contain noble tetracyclic triterpenoid saponins derived from dammarenediol-II. Dammarene-type ginsenosides are classified into the protopanaxadiol (PPD) and protopanaxatriol (PPT) groups based on their triterpene aglycone structures. Two cytochrome P450 (CYP) genes (CYP716A47 and CYP716A53v2) are critical for the production of PPD and PPT aglycones, respectively. CYP716A53v2 is a protopanaxadiol 6-hydroxylase that catalyzes PPT production from PPD in P. ginseng. Methods: We constructed transgenic P. ginseng lines overexpressing or silencing (via RNA interference) the CYP716A53v2 gene and analyzed changes in their ginsenoside profiles. Result: Overexpression of CYP716A53v2 led to increased accumulation of CYP716A53v2 mRNA in all transgenic roots compared to nontransgenic roots. Conversely, silencing of CYP716A53v2 mRNA in RNAi transgenic roots resulted in reduced CYP716A53v2 transcription. HPLC analysis revealed that transgenic roots overexpressing CYP716A53v2 contained higher levels of PPT-group ginsenosides ($Rg_1$, Re, and Rf) but lower levels of PPD-group ginsenosides (Rb1, Rc, $Rb_2$, and Rd). By contrast, RNAi transgenic roots contained lower levels of PPT-group compounds and higher levels of PPD-group compounds. Conclusion: The production of PPD- and PPT-group ginsenosides can be altered by changing the expression of CYP716A53v2 in transgenic P. ginseng. The biological activities of PPD-group ginsenosides are known to differ from those of the PPT group. Thus, increasing or decreasing the levels of PPT-group ginsenosides in transgenic P. ginseng may yield new medicinal uses for transgenic P. ginseng.

• Journal of Microbiology and Biotechnology
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• 제21권10호
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• pp.1057-1063
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• 2011

Herein, a novel ginsenosidase, named ginsenosidase type IV, hydrolyzing 6-O-multi-glycosides of protopanaxatriol-type ginsenosides (PPT), such as Re, R1, Rf, and Rg2, was isolated from the Aspergillus sp. 39g strain, purified, and characterized. Ginsenosidase type IV was able to hydrolyze the 6-O-${\alpha}$-L-($1{\rightarrow}2$)-rhamnoside of Re and the 6-O-${\beta}$-D-($1{\rightarrow}2$)-xyloside of R1 into ginsenoside Rg1. Subsequently, it could hydrolyze the 6-O-${\beta}$-D-glucoside of Rg1 into F1. Similarly, it was able to hydrolyze the 6-O-$_{\alpha}$-L-($1{\rightarrow}2$)-rhamnoside of Rg2 and the 6-O-${\beta}$-D-($1{\rightarrow}2$)-glucoside of Rf into Rh1, and then further hydrolyze Rh1 into its aglycone. However, ginsenosidase type IV could not hydrolyze the 3-O- or 20-O-glycosides of protopanaxadiol-type ginsenosides (PPD), such as Rb1, Rb2, Rb3, Rc, and Rd. These exhibited properties are significantly different from those of glycosidases described in Enzyme Nomenclature by the NC-IUBMB. The optimal temperature and pH for ginsenosidase type IV were $40^{\circ}C$ and 6.0, respectively. The activity of ginsenosidase type IV was slightly improved by the $Mg^{2+}$ ion, and inhibited by $Cu^{2+}$ and $Fe^{2+}$ ions. The molecular mass of the enzyme, based on SDS-PAGE, was noted as being approximately 56 kDa.

• Journal of Microbiology and Biotechnology
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• 제23권4호
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• pp.444-450
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• 2013

A Gram-negative, strictly aerobic, non-motile, non-spore-forming, and rod-shaped bacterial strain designated FW-$6^T$ was isolated from a freshwater sample and its taxonomic position was investigated by using a polyphasic approach. Strain FW-$6^T$ grew optimally at $10-42^{\circ}C$ and at pH 7.0 on nutrient and R2A agar. Strain FW-$6^T$ displayed ${\beta}$-glucosidase activity that was responsible for its ability to transform ginsenoside $Rb_1$ (one of the dominant active components of ginseng) to Rd. On the basis of 16S rRNA gene sequence similarity, strain FW-$6^T$ was shown to belong to the family Sphingomonadaceae and was related to Novosphingobium aromaticivorans DSM $12444^T$ (98.1% sequence similarity) and N. subterraneum IFO $16086^T$ (98.0%). The G+C content of the genomic DNA was 64.4%. The major menaquinone was Q-10 and the major fatty acids were summed feature 7 (comprising $C_{18:1}{\omega}9c/{\omega}12t/{\omega}7c$), summed feature 4 (comprising $C_{16:1}{\omega}7c/iso-C_{15:0}2OH$), $C_{16:0}$, and $C_{14:0}$ 2OH. DNA and chemotaxonomic data supported the affiliation of strain FW-$6^T$ to the genus Novosphingobium. Strain FW-$6^T$ could be differentiated genotypically and phenotypically from the recognized species of the genus Novosphingobium. The isolate that has ginsenoside converting ability therefore represents a novel species, for which the name Novosphingobium ginsenosidimutans sp. nov. is proposed, with the type strain FW-$6^T$ (= KACC $16615^T$ = JCM $18202^T$).