• Journal of Microbiology and Biotechnology
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• 제28권2호
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• pp.255-261
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• 2018

Aglycon protopanaxatriol (APPT) has valuable pharmacological effects such as memory enhancement and tumor inhibition. ${\beta}$-Glycosidase from the hyperthermophilic bacterium Dictyoglomus turgidum (DT-bgl) hydrolyzes the glucose residues linked to APPT, but not other glycoside residues. ${\beta}$-Glycosidase from the hyperthermophilic bacterium Pyrococcus furiosus (PF-bgl) hydrolyzes the outer sugar at C-6 but not the inner glucose at C-6 or the glucose at C-20. Thus, the combined use of DT-bgl and PF-bgl is expected to increase the biotransformation of PPT-type ginsenosides to APPT. We optimized the ratio of PF-bgl to DT-bgl, the concentrations of substrate and enzyme, and the reaction time to increase the biotransformation of ginsenoside Re and PPT-type ginsenosides in Panax ginseng leaf extract to APPT. DT-bgl combined with PF-bgl converted 1.0 mg/ml PPT-type ginsenosides in ginseng leaf extract to 0.58 mg/ml APPT without other ginsenosides, with a molar conversion of 100%. We achieved the complete biotransformation of ginsenoside Re and PPT-type ginsenosides in ginseng leaf extract to APPT by the combined use of two ${\beta}$-glycosidases, suggesting that discarded ginseng leaves can be used as a source of the valuable ginsenoside APPT. To the best of our knowledge, this is the first quantitative production of APPT using ginsenoside Re, and we report the highest concentration and productivity of APPT from ginseng extract to date.

• Journal of Ginseng Research
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• 제42권4호
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• pp.412-418
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• 2018

Background: Ginsenoside Rg3(S) and compound K (C-K) are pharmacologically active components of ginseng that promote human health and improve quality of life. The aim of this study was to produce Rg3(S) and C-K from ginseng extract using recombinant Lactococcus lactis. Methods: L. lactis subsp. cremoris NZ9000 (L. lactis NZ9000), which harbors ${\beta}$-glucosidase genes (BglPm and BglBX10) from Paenibacillus mucilaginosus and Flavobacterium johnsoniae, respectively, was reacted with ginseng extract (protopanaxadiol-type ginsenoside mixture). Results: Crude enzyme activity of BglBX10 values comprised 0.001 unit/mL and 0.003 unit/mL in uninduced and induced preparations, respectively. When whole cells of L. lactis harboring pNZBglBX10 were treated with ginseng extract, after permeabilization of cells by xylene, Rb1 and Rd were converted into Rg3(S) with a conversion yield of 61%. C-K was also produced by sequential reactions of the permeabilized cells harboring each pNZBgl and pNZBglBX10, resulting in a 70% maximum conversion yield. Conclusion: This study demonstrates that the lactic acid bacteria having specific ${\beta}$-glucosidase activity can be used to enhance the health benefits of Panax ginseng in either fermented foods or bioconversion processes.

• 한국약용작물학회지
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• 제20권4호
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• pp.270-276
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• 2012

This study was performed to enhance contents of low molecular weight ginsenoside Rh2 and Rg3 using an ultra high pressure and steaming process in wild cultured-Root in wild ginseng. For selective increase in contents of Rg3 and Rh2 in cultured wild ginseng roots, an ultra high extraction was applied at 500MPa for 20 min which was followed by steaming process at $90^{\circ}C$ for 12 hr. It was revealed that contents of ginsenosides, Rb1, Rb2, Rc and Rd, were decreased with the complex process described above, whereas contents of ginsenoside Rh2 and Rg3 were increased up to 4.918 mg/g and 6.115 mg/g, respectively. In addition, concentration of benzo[${\alpha}$]pyrene in extracts of the cultured wild ginseng roots treated by the complex process was 0.64 ppm but it was 0.78 ppm when it was treated with the steaming process. From the results, it was strongly suggested that low molecular weight ginsenosides, Rh2 and Rg3, are converted from Rb1, Rb2, Rc, and Rd which are easily broken down by an ultra high pressure and steaming process. This results indicate that an ultra high pressure and steaming process can selectively increase in contents of Rg3 and Rh2 in cultured wild ginseng roots and this process might enhance the utilization and values of cultured wild ginseng roots.

• 미생물학회지
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• 제54권4호
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• pp.436-441
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• 2018

인삼은 우수한 약리 활성 작용을 보이는 전통적인 약초이다. 본 연구에는 프로바이오틱스 Lactobacillus helveticus KII13과 Pediococcus pentosaceus KID7 균주를 진세노사이드(ginsenoside) 함량을 증가시키기 위해 조 인삼 추출물을 발효시켜 진세노사이드를 형질전환 시키는데 사용되었다. 발효삼 추출물의 TLC(Thin-layer chromatography) 분석 결과, 5일간의 발효 후 주요 사포닌인 진세노사이드 Rg3, Rh1 및 Rh2로 변환되는 것으로 나타났다. HPLC 분석을 수행하여 주요 및 미량 진 세노사이드를 정량화하였다. 3일째에는 Rg3가 나타나고, 5일째에는 Rh2 및 Rh1이 나타난다. L. helveticus KII13과 P. pentosaceus KID7의 공동 배양은 주요 진세노사이드(Rb1과 Rg1)를 미량 진세노사이드(Rg3, Rh2, Rh1)로 전환시키는 것을 학인하였다.

• The Korean Journal of Physiology and Pharmacology
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• 제17권2호
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• pp.127-132
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• 2013

Ginsenosides, one of the active ingredients of Panax ginseng, show various pharmacological and physiological effects, and they are converted into compound K (CK) or protopanaxatriol (M4) by intestinal microorganisms. CK is a metabolite derived from protopanaxadiol (PD) ginsenosides, whereas M4 is a metabolite derived from protopanaxatriol (PT) ginsenosides. The ${\gamma}$-aminobutyric acid $receptor_C$ ($GABA_C$) is primarily expressed in retinal bipolar cells and several regions of the brain. However, little is known of the effects of ginsenoside metabolites on $GABA_C$ receptor channel activity. In the present study, we examined the effects of CK and M4 on the activity of human recombinant $GABA_C$ receptor (${\rho}$ 1) channels expressed in Xenopus oocytes by using a 2-electrode voltage clamp technique. In oocytes expressing $GABA_C$ receptor cRNA, we found that CK or M4 alone had no effect in oocytes. However, co-application of either CK or M4 with GABA inhibited the GABA-induced inward peak current ($I_{GABA}$). Interestingly, pre-application of M4 inhibited $I_{GABA}$ more potently than CK in a dose- dependent and reversible manner. The half-inhibitory concentration ($IC_{50}$) values of CK and M4 were $52.1{\pm}2.3$ and $45.7{\pm}3.9{\mu}M$, respectively. Inhibition of $I_{GABA}$ by CK and M4 was voltage-independent and non-competitive. This study implies that ginsenoside metabolites may regulate $GABA_C$ receptor channel activity in the brain, including in the eyes.

• Journal of Ginseng Research
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• 제42권4호
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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.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 2002년도 학술대회지
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• pp.486-490
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• 2002

In different approaches ginsenosides were extracted from Korean ginseng roots by ammonia and for comparison with methanol-water and water. The extracts have been analyzed qualitatively and quantitatively to evaluate yield and selectivity of extractions of ginsenosides. Water supplied the lowest yield. The yields of extracts with liquid ammonia were higher than those with methanol-water. However, this is partly due to the conversion of malonyl ginsenoside to normal ginsenosides by ammonia. It was proved by HPLC that malonyl-ginsenosides $m-Rb_1,\;m-Rb_2,$ m-Rc and m-Rd were converted to the corresponding neutral ginsenosides. Furthermore, ginsenosides from ginseng roots were extracted by alkaline methanol-water $(60\%)$ solutions. Alternatively, the extracts of the methanol-water $(60\%)$ extraction were treated with sodium hydroxide solution. Both methods also convert the malonyl-ginsenosides to neutral ginsenosides.

• 한국자원식물학회지
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• 제25권4호
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• pp.473-481
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• 2012

구증구포방법은 기존의 홍삼제조방법에서와 같이 9회 반복 과정으로 새로운 신규사포닌 등 성분변화가 일어나지만 시간이 오래 걸리고 복잡하며 어떤 특수 성분이 얼마나 증가 되는지 보고 되어 있지 않다. 또한 기존의 구증구포방법은 제조공정 중 건조시 보통 $60^{\circ}C$에서 열풍건조를 하기 때문에 건조시 관리의 부족으로 간혹 벤조피렌에 노출되는 경우가 있다. 본 방법은 새로운 자동 구증구포방법으로 제조시간이 약 2배정도 단축되며 특히 건조시 습열냉각건조를 통하기 때문에 벤조피렌함량이 거의 검출되지 않았다. 또한 사포닌 변환 등은 기존 구증구포방법과 같이 사포닌 변화가 일어나 홍삼에서만 나타나는 Rg3와 기타 효능활성물질 등이 분석되었다. 인삼사포닌의 경우에는 증포횟수가 증가함에 따라 흡수가 어려운 major ginsenoside(Rg1, Re, Rb1, Rc, Rb2 및 Rd)의 함량이 점차적으로 감소되고 대신 흡수가 빠르고 항암활성이 강한 minor ginsenoside (Rh1, 20(S)-Rg2, 20(R)-Rg2, 20(S)-Rg3, 20(R)-Rg3, Rk1 및 Rg5)의 함량이 점차적으로 증가하였다. 특히 diol계 사포닌인 ginsenosides Rb1, Rb2, Rc 및 Rd는 Rg3, Rk1 및 Rg5로 전환되었고, triol계 사포닌인 ginsenosides Rg1 및 Re는 Rh1, Rg2로 전환되었다. 수삼에서의 환원당, 산성다당체 및 총 페놀 화합물 함량은 7회까지 유의적으로 증가하였고 8회부터 점차 감소하는 경향을 보였다. DPPH 라디칼 소거활성은 7회까지 점차적으로 감소하여 $IC_{50}$값이 68% 감소되는 것으로 나타났으며 7회부터 9회까지는 큰 유의적 차이가 없었다. 결론적으로 본 자동 구중구포방법은 기존의 방법과 물질생성은 거의 비슷하지만 시간이 단축되고 벤조피렌 함량이 거의 검출되지 않아 앞으로 고부가가치 인삼산업에 많은 도움을 줄 것으로 생각된다.

• Journal of Ginseng Research
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• 제39권4호
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• pp.384-391
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• 2015

It has been reported that Korean Red Ginseng has been manufactured for 1,123 y as described in the GoRyeoDoGyeong record. The Korean Red Ginseng manufactured by the traditional preparation method has its own chemical component characteristics. The ginsenoside content of the red ginseng is shown as Rg1: 3.3 mg/g, Re: 2.0 mg/g, Rb1: 5.8 mg/g, Rc:1.7 mg/g, Rb2: 2.3 mg/g, and Rd: 0.4 mg/g, respectively. It is known that Korean ginseng generally consists of the main root and the lateral or fine roots at a ratio of about 75:25. Therefore, the red ginseng extract is prepared by using this same ratio of the main root and lateral or fine roots and processed by the historical traditional medicine prescription. The red ginseng extract is prepared through a water extraction ($90^{\circ}C$ for 14-16 h) and concentration process (until its final concentration is 70-73 Brix at $50-60^{\circ}C$). The ginsenoside contents of the red ginseng extract are shown as Rg1: 1.3 mg/g, Re: 1.3 mg/g, Rb1: 6.4 mg/g, Rc:2.5 mg/g, Rb2: 2.3 mg/g, and Rd: 0.9 mg/g, respectively. Arginine-fructose-glucose (AFG) is a specific amino-sugar that can be produced by chemical reaction of the process when the fresh ginseng is converted to red ginseng. The content of AFG is 1.0-1.5% in red ginseng. Acidic polysaccharide, which has been known as an immune activator, is at levels of 4.5-7.5% in red ginseng. Therefore, we recommended that the chemical profiles of Korean Red Ginseng made through the defined traditional method should be well preserved and it has had its own chemical characteristics since its traditional development.

• 한국미생물·생명공학회지
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• 제50권3호
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• pp.414-421
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• 2022

식물 공생 미생물은 기주 식물과 함께 공생하는 미생물로 생장 촉진, 면역력 증진, 대사체 생성 등의 역할을 수행하며 식물 발달에 영향을 준다. 본 연구를 통해 30년근 산삼에서 분리 동정한 미생물인 W. ginsengihumi LGHNH (KCTC 14462BP)은 식물 생장 촉진 호르몬인 indole-3-acetic acid (IAA)을 1.38 ㎍/ml에서 2.22 ㎍/ml 수준으로 분비함을 확인하였다. 또한 발효 전, 후의 진세노사이드 함량 비교를 통해 진세노사이드 전환능이 있음을 확인하였다. 전환된 저분자 진세노사이드인 Rg2(R), Rg4, Rg6, Rg3(S), Rg3(R), Rk1, Rg5, Rh1(R), Rk3, Rh4 등은 생체 이용률이 높고 다양한 피부 효능을 갖는다고 알려져 있다. 배양물로 제조한 W. ginsengihumi LGHNH (W. ginsengihumi LGHNH Cultured product, WCP)의 항노화 소재로서 가능성을 탐색하기 위해 미토콘드리아의 막전위와 ATP 생합성량을 측정하여 기능 저하 억제 여부를 확인하였다. 노화를 발생시키는 인자인 UVB를 조사한 HaCaT 세포 내 미토콘드리아 막전위 값을 측정한 결과, 미조사군 대비 39.3%로 감소하나 WCP 0.001% (v/v), 0.01% (v/v)에 의해 각각 57.3%, 58.1% 수준까지 회복함을 확인하였다. 또한 미토콘드리아의 ATP 생합성량 측정 결과, UVB 조사에 의해 미조사군 대비 94.3% 수준으로 감소하나 WCP를 0.001% (v/v), 0.01% (v/v) 처리한 군에서 각 각 105.3%, 105.7%로 증가하여 미토콘드리아 기능을 정상으로 회복하는데 도움을 줄 수 있다고 판단된다. 따라서, 본 연구를 통해 확보한 30년근 산삼의 공생 미생물은 항노화 관련 생물 자원으로서 산업적 활용 가능성이 높다.