• Journal of Ginseng Research
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• 제27권2호
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• pp.78-85
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• 2003

Agrobacterium rhizognes $A_4$ Ti-plasmid를 인삼 뿌리에 도입하여 유전적으로 안정하며 반영구적으로 이용할 수 있는 형질전환된 인삼모상근 캘러스(GHC-T78)를 유도하였다. 특정 ginsenoside를 대량으로 생산할 수 있는 최적배양조건을 확립하기 위해서 먼저 모상근으로부터 캘러스의 유도하는 과정에서 BA의 단독 처리구는 auxin과 cytokinin의 혼합 처리구보다 캘러스 형성이 더 우수하였다. 인삼 캘러스 형성율은 Benzyladenine(BA)를 1 mg-3 mg/L 까지 첨가한 배지에서 가장 높게 나타났으며, ginsenoside 함량 역시 bezyladenine 단독처리구는 가장 우수하였다. 캘러스 세포의 액체배양 기간을 4주로 설정할 때 BA의 농도 역시 BA 2 mg/L로 첨가한 배지에서 가장 우수한 캘러스 생장률을 보였다. 실제로 인삼 모상근 캘러스의 생장율은 암상태에서 배양하는 것이 광상태에서 보다 다소 양호하였으나, ginsenoside의 생성에는 연속광상태에서 더 효과적임이 확인되었다. 모상근 캘러스에서 얻은 이러한 결과는 국내외 처음으로 보고되는 내용이며, 액체배양의 캘러스 세포에서 분화된 무수히 많은 단일 모상근 근모의 형성은 액체배양을 통한 특정 ginsenosid의 새로운 생산방법을 개발할 수 있도록 하고 있다.

• 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.

• Applied Biological Chemistry
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• 제51권2호
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• pp.114-118
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• 2008

Ginseng treated with several treatment conditions of various acids to search hydrolysates on the basis of increased biological activity and modified structure. In the result of acid treatment, the conversion rate of ginsenoside Rg3, Rk1 and Rg5 was highest when ginseng treated with citric acid. After added citric acid to ginseng extract, boiled at l00$^{\circ}C$ for 1 hour and add enzyme, which is examined change by time. It compared with group which did not treated acid. Two groups became difference according to enzyme but the generation rate of ginsenoside Rg3, Rk1 and Rg5 did not show difference greatly. Also, the generation rate of ginsenoside Rg3, Rk1 and Rg5 by time passes did not show difference. The generation rate of ginsenoside Rg3, Rk1 and Rg5 increased when increased acid concentration, temperature and time. We did exclusion molding to shorten treatment time. In the result of ginseng treated with citric acid of various concentrations at various temperatures as time passes by extrusion molding, the generation rate of ginsenoside Rg3, Rk1 and Rg5 was highest when ginseng treated with 3% citric acid at l60$^{\circ}C$ for 20 minutes. In addition, total saponin amount of ginseng treated with 3% citric acid at 160$^{\circ}C$ for 20 minutes was about 11% higher than ginseng heated at 120$^{\circ}C$ for 3 hours. These results indicated that our exclusion molding process more effective, compared to traditional red ginseng manufacturing process.

• Archives of Pharmacal Research
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• 제29권2호
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• pp.145-151
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• 2006

We investigated the effects of Ginsenoside $R_e$ on human sperm motility in fertile and asthenozoospermic infertile individuals in vitro and the mechanism by which the Ginsenosides play their roles. The semen samples were obtained from 10 fertile volunteers and 10 asthenozoospermic infertile patients. Spermatozoa were separated by Percoll and incubated with 0, 1, 10 or $100\;{\mu}M$ of Ginsenoside $R_e$. Total sperm motility and progressive motility were measured by computer-aided sperm analyzer (CASA). Nitric oxide synthase (NOS) activity was determined by the $^{3}H$-arginine to $^{3}H$-citrulline conversion assay, and the NOS protein was examined by the Western blot analysis. The production of sperm nitric oxide (NO) was detected using the Griess reaction. The results showed that Ginsenoside $R_e$ significantly enhanced both fertile and infertile sperm motility, NOS activity and NO production in a concentration-dependent manner. Sodium nitroprusside (SNP, 100 nM), a NO donor, mimicked the effects of Ginsenoside $R_e$. And pretreatment with a NOS inhibitor $N^{w}$-Nitro-L-arginine methyl ester (L-NAME, $100\;{\mu}M$) or a NO scavenger N-Acetyl-L-cysteine (LNAC, 1 mM) completely blocked the effects of Ginsenoside $R_e$. Data suggested that Ginsenoside $R_e$ is beneficial to sperm motility, and that induction of NOS to increase NO production may be involved in this benefit.

• Journal of Microbiology and Biotechnology
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• 제29권3호
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• pp.410-418
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• 2019

To investigate a novel ${\beta}$-glucosidase from Bifidobacterium breve ATCC 15700 (BbBgl) to produce compound K (CK) via ginsenoside $F_2$ by highly selective and efficient hydrolysis of the C-3 glycoside from ginsenoside Rd, the BbBgl gene was cloned and expressed in E. coli BL21. The recombinant BbBgl was purified by Ni-NTA magnetic beads to obtain an enzyme with specific activity of 37 U/mg protein using pNP-Glc as substrate. The enzyme activity was optimized at pH 5.0, $35^{\circ}C$, 2 or 6 U/ml, and its activity was enhanced by $Mn^{2+}$ significantly. Under the optimal conditions, the half-life of the BbBgl is 180 h, much longer than the characterized ${\beta}$-glycosidases, and the $K_m$ and $V_{max}$ values are 2.7 mM and $39.8{\mu}mol/mg/min$ for ginsenoside Rd. Moreover, the enzyme exhibits strong tolerance against high substrate concentration (up to 40 g/l ginsenoside Rd) with a molar biotransformation rate of 96% within 12 h. The good enzymatic properties and gram-scale conversion capacity of BbBgl provide an attractive method for large-scale production of rare ginsenoside CK using a single enzyme or a combination of enzymes.

• 한국식품영양학회지
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• 제33권4호
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• pp.372-380
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• 2020

The purpose of this study is to investigate the physicochemical properties of jelly made from fermented red ginseng concentrate (FRGC) that can be easily absorbed and digested for the health promotion of the elderly. The pH of the jellies tended to decrease with increasing concentration of FRGC. Soluble solid content has significantly higher value when added more than 2%, and the water content of the sample was significantly lower when the FRGC was added 4%. As the amount of FRGC was increased, the total color difference increased, and the hardness of samples decreased significantly. On the other hand, the total ginsenoside contents of the FRGC was 45.50 mg/g. As the concentration of FRGC increased, the content of polyphenol and flavonoids increased. The increasing pattern of polyphenols and flavonoids showed a similar trend. As the content of FRGC increased, ABTS free radical scavenging activity significantly increased (p<0.05), and in the control, the minimum value (62.6 AEAC) and the 4% sample were highest (116.2 AEAC). DPPH radical scavenging activity was like that of ABTS radical scavenging activity. However, there was no significant difference in DPPH radical scavenging activity of 3% and 4% red ginseng jelly.

• Food Science and Biotechnology
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• 제16권5호
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• pp.848-853
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• 2007

The effects of high-hydrostatic pressure (HHP) on the ginsenoside concentration in Korean red ginseng were investigated. HHP-pretreated Korean red ginseng samples were compared to samples produced by a conventional method. Six-year-old Korean fresh ginseng (Panax ginseng C.A. Meyer) samples were vacuum-packaged in polyethylene film and treated at room temperature for 1 min with HHP (200-600 MPa) and steamed at $98^{\circ}C$ for 3 hr. Major ginsenosides of red ginseng were analyzed by HPLC. HHP-pretreated red ginseng showed a 45% higher level of total major ginsenosides than conventionally prepared red ginseng. The levels of 4 protopanaxadiol-type ginsenosides increased 34-43% and the levels of 5 protopanaxatriol-type ginsenosides increased 45-66%. Scanning electron microscopy and electrical conductivity spectrum analysis showed that HHP pretreatment damaged ginseng plant cells and increased extraction efficiencies of ginsenosides from red ginseng products.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 1998년도 Advances in Ginseng Research - Proceedings of the 7th International Symposium on Ginseng -
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• pp.182-189
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• 1998

Administration of ginsenosides, a mixture of saponin extracted from Panax ginseng, decreased blood pressure in rat. Previous studies have shown that ginsenosides caused endothelium-dependent relaxation, which was associated with the formation of cyclic GMP, suggested that ginsenosides caused release of nitric oxide (NO) from the vascular endothelium. The aim of the present study was to characterize the endothelium-independent relaxation to ginsenosides in the isolated rat aorta. Ginsenosides caused a concentration-dependent relaxation of rat aortic rings without endothelium constricted with 25 mM KCI but affected only minimally those constricted with 60 mM KCI. Ginsenoside Rg3 (Rg3) was a more potent vasorelaxing agonist than total ginsenoside mixture and also the ginsenoside PPT and PPD groups. Relaxation to ginsenosides were markedly reduced by TEA, but not by glibenclamide. Rg3 significantly inhibited Cal'-induced concentration-contraction curves and the "50a2'influx in aortic rings incubated in 25 mM KCI whereas those responses were not affected in 60 mM KCI. Rg3 caused efflux of $"Rb in aortic rings that was inhibited by tetraethy- lammonium (TEA), an inhibitor of Ca"-dependent K'channels, but not by glibenclamide, an inhibitor of AfP-dependent K'channels. These findings indicate that ginsenosides may induce vasorelaxation via activation of Ca2'-dependent K'channels resulting in hyperpolarization of the vas- cular smooth muscle with subsequent inhibition of the opening of voltage-dependent Caf'channels. These effects could contribute to explain the red ginseng-associated vasodilation and the beneficial effect on the cardiovascular system.

• 한국동물위생학회지
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• 제41권3호
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• pp.141-147
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• 2018

Red ginseng marc is a by-product of Korean red ginseng (panax ginseng CA Meyer) and contains ginsenoside which has pharmacological effects. The Korean red ginseng marc was fermented with Bacillus subtilis (RGMB). This study was carried out to investigate the RGMB effect on swine immunity. The variation of ginsenoside depending on the RGMB fermentation time was analyzed. Swine (Landrace${\times}$Yorkshire) were divided into control group (basic diet) and RGMB group (RGMB 1% diet). One percent RGMB was fed to the RGMB group for 28 days. The biochemical parameters, cytokine and immunoglobulin were analyzed. For 48 hours of fermentation on RGMB, ginsenoside Rb1 had increased 180.94%, Rg3 235.85%. Rg1 wasn't detected before fermentation, but was detected after 48 hours of fermentation. The RGMB had effect of deceasing initial AST concentration $79.33{\pm}12.85U/L$ to $54.00{\pm}14.46U/L$ in final and was significantly lower (P<0.05) than control in final. In final RGMB had significantly lower (P<0.05) ALT concentration of $48.57{\pm}8.26U/L$ comparing with control group of $65.43{\pm}10.31U/L$. RGMB had the effect of significantly decreasing (P<0.05) $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ concentration of $2.44{\pm}1.31ng/mL$, $0.71{\pm}0.36ng/mL$ and $0.51{\pm}0.21ng/mL$. The IgA concentration had significantly increased (P<0.05) in RGMB group of $0.56{\pm}0.06mg/mL$ in final. These results demonstrate that RGMB has effect of increasing immunity and practicable to use as feed additives on swine.

• 동아시아식생활학회지
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• 제7권4호
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• pp.484-490
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• 1997

백삼성분이 탐식세포를 통한 면역반응조절에 어떠한 영향을 미치는가를 알아보기 위해 total saponin이나 ginsenoside Rb$_2$성분이 마우스 복강 탐식 세포의 증식과 nitric oxide(NO)분비 및 NOS 유전자 발현에 미치는 영향을 조사해 보았다. Total saponin 또는 ginsenoside Rb$_2$성분을 탐식세포 배양액에 0-256$\mu\textrm{g}$/$m\ell$ 농도로 첨가하여 배양한 후 $^3$H-thymidine incorporation법으로 분석을 실시한 결과 total saponin은 64$\mu\textrm{g}$/$m\ell$ 농도에서 세포내 DNA 합성을 증가시켰으며, ginsenoside Rb$_2$ 성분의 경우 16$\mu\textrm{g}$/$m\ell$ 또는 64$\mu\textrm{g}$/$m\ell$ 농도에서 DNA 합성이 증가되었다. 256$\mu\textrm{g}$/$m\ell$의 높은 농도에서 세포의 증식이 약간 저해되었지만 대조군에 비해 유의적인 감소는 보이지 않았다. NO 분비에 미치는 영향을 조사해 본 결과 20$\mu\textrm{g}$/$m\ell$ 농도의 백삼성분을 투여했을 때 NO분비가 대조군에 비해 유의적으로 증가하였다. Reverse transcription polymerase chain reaction (RT-PCR)을 이용하여 백삼성분에 의한NOS 유전자 발현의 정도를 조사한 결과 20$\mu\textrm{g}$/$m\ell$ 농도에서 nos gene발현이 대조군보다 증가되었다