• 대한화장품학회지
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• 제49권4호
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• pp.375-383
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• 2023

최근 다양한 피부 기능 개선 효과로 기능성 소재로서 활용도가 높은 홍삼 사포닌의 한 종류인 진세노사이드 Rg3를 위한 사과산(malic acid)활용 전환 방법을 확인하였다. 실험 계획법인 반응 표면 분석법(RSM)을 활용하여 진세노사이드 Rg3로의 전환에 영향을 주는 요인을 최적화하기 위한 실험 조건을 설계 및 검증하였다. 주요 독립변수는 사과산 농도, 반응 온도와 반응 시간이었고 Box-Behnken design (BBD)법에 따라 설계된 실험 조건으로 진세노사이드 Rg3로 전환을 수행하고 최적화 조건을 분석하였다. 전환된 진세노사이드 Rg3의 농도는 1.548 mg/L에서 최대 4.558 mg/L까지 확인되었고 사과산 1%, 50℃, 9 h에서 가장 높은 양의 진세노사이드 Rg3생성량을 보였다. 결론적으로, 진세노사이드 Rg3의 생성에 가장 영향을 미치는 요인은 사과산의 농도, 반응 시간, 온도 순이었다. 또한, 사과산의 농도와 반응 시간의 교호작용이 반응 온도 요인보다 영향도가 큰 것을 확인하였다.

• 한국축산식품학회지
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• 제37권5호
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• pp.735-742
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• 2017

This study was conducted to isolate and characterize Paenibacillus sp. MBT213 possessing ${\beta}$-glucosidase activity from raw milk, and examine the enzymatic capacity on the hydrolysis of a major ginsenoside ($Rb_1$). Strain MBT213 was found to have a high hydrolytic ability on ginsenoside $Rb_1$ by Esculin Iron Agar test. 16S rDNA analysis revealed that MBT213 was Paenibacillu sp. Crude enzyme of MBT213 strain exhibited high conversion capacity on ginsenoside $Rb_1$ into ginsenoside Rd proven by TLC and HPLC analyses. The API ZYM kit confirmed that Paenibacillu sp. MBT213 exerted higher ${\beta}$-glucosidase and ${\beta}$-galactosidase activity than other strains. Optimum pH and temperature for crude enzyme were found at 7.0 and $35^{\circ}C$ in hydrolysis of ginsenoside $Rb_1$. After 10 d of optimal reaction conditions for the crude enzyme, ginsenoside $Rb_1$ fully converted to ginsenoside Rd. Ginseng roots (20%) were fermented for 14 d, and analyzed by HPLC showed that amount of ginsenoside $Rb_1$ significantly decreased, while that of ginsenoside Rd was significantly increased. The study confirmed that the ${\beta}$-glucosidase produced by Paenibacillus sp. MBT213 can hydrolyze the major ginsenoside $Rb_1$ and convert to Rd during fermentation of the ginseng. The ${\beta}$-glucosidase activity of this novel Paenibacillus sp. MBT213 strain may be utilized in development of variety of health foods, dairy foods and pharmaceutical products.

• 대한화장품학회지
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• 제46권4호
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• pp.349-360
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• 2020

최근 피부 기능 개선과 관련한 다양한 가능성으로 인해 화장품 소재로서 수요가 높아지고 있는 인삼 유래 사포닌의 한 종류인 ginsenoside Rd를 위한 생물 전환 제조 기술을 확립하였다. 홍삼 사포닌(RGS)에 포함된 ginsenoside Rb1을 Rd로 전환하기 위하여 상업용 효소를 탐색하였고 그 중 Viscoflow MG가 가장 효율적인 것을 확인하였다. Ginsenoside Rd로의 전환에 영향을 주는 요인을 최적화하기 위하여 반응표면분석법(RSM)을 통하여 실험 조건을 설계하였다. 주요 독립변수는 RGS 농도, 효소 농도와 반응 시간이었고 Box-Behnken design (BBD) 모델설계법에 따라 선정된 17 가지 조건으로 ginsenoside Rd로 전환을 수행하고 최적화 조건을 분석하였다. 전환된 Ginsenoside Rd의 농도는 0.3113 g/L에서 최대 0.5277 g/L까지였고 RGS 2%, 효소 1.25%를 13.5 h 반응시킨 조건에서 가장 높은 생성량을 보였다. 결론적으로, ginsenoside Rd 생물전환의 독립변수인 RGS 농도, 효소 농도는 p-value가 0.05보다 작은 값으로 유의미한 값을 나타내었고 각 독립변수 사이의 교호작용 중에서는 효소 농도와 반응 시간 사이의 교호 작용이 가장 큰 영향력을 갖고 있음을 확인하였다.

• 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.

• 대한화장품학회지
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• 제41권4호
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• pp.375-382
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• 2015

진세노사이드(인삼 사포닌)는 인삼의 대표적 약리성분 중의 하나로 생물학적 활성을 가진 배당체 화합물이다. 이들 사포닌은 가수분해 되어 저분자화 되었을 때, 항주름 및 항산화, 항암 등에 높은 약리효능효과를 나타낸다. 본 연구에서는 인삼 esculin 배지를 활용하여 ${\beta}$-glucosidase 활성을 가진 균주를 분리하였고 인삼 사포닌 전환을 미생물을 이용하여 수행하였다. 본 균주들을 16S rRNA sequencing을 통하여 동정하여 본 결과 Stenotrophomonas rhizopilae strain GFC09로 확인되였다. 균주의 최적 활성 조건을 결정하기 위해 조효소 1 mM와 인삼사포닌 $Rb_1$과 함께 배양한 후 생물학적 전환을 TLC, HPLC를 사용하여 확인하였다. 조효소에 의한 인삼 사포닌 $Rb_1$의 전환 경로는 다음과 같다. LB: RbNeobio R&D center, Gyeonggi-do 16954, Korea${\rightarrow}$Rd${\rightarrow}$FNeobio R&D center, Gyeonggi-do 16954, Korea${\rightarrow}$compound K, TSB: $Rb_1{\rightarrow}Rd{\rightarrow}F_2$. 가수분해된 생성된 물질은 NMR로 구조 동정하였다. 전환 산물의 효능 분석결과, 콜라겐 생성을 농도 의존적으로 증가시키는 것이 관찰되었다. 이에 본 연구에서는 ginsenoside $F_2$와 compound K 함유 인삼 전환 산물의 주름 개선 소재로서 활용가능성을 확인하였다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2010년도 정기총회 및 추계학술발표회
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• pp.20-20
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• 2010

Panax ginseng C.A Meyer is frequently taken orally as a traditional herbal medicine in Asian countries. The major components of ginseng are ginsenoside, which are pharmaceutical activity. The six major ginsenosides, including Rb1, Rb2, Rc, Rd, Re and Rg1 account for 90% of total ginsenosides. Even though the minor ginsenosides, including Rg3, Rh2 and compound K has high pharmacetical activities, the price of minor ginsenosides is too high. Therefore we isolated the gypenoside V and made it converted to minor ginsenosides. In the plant Gynostemma pentaphyllum Makino, gypenosdie V was presented as dominant saponin (content about 2.4%), and was similar to protopanaxadol type ginsenosides such as ginsenoside Rb1. In this study, we confirmed that the coversion of gypenoside V to minor ginsenosides after using the various treatment such as heating, acid treatment, commercial edible enzyme, and lactobacillus. Consequently, we optimizied the transformation of gypenoside V to minor ginsenoside using Thin Layer Chromatography (TLC), High Performance Liquid Chromatography (HPLC), Time-of-flight Mass Spectrometry (LC/TOF/MS).

• 대한약침학회지
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• 제23권2호
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• pp.79-87
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• 2020

Objectives: Ginsenosides found in ginseng, and the hydrolysates derived from their conversion, exhibit diverse pharmacological characteristics [1]. These have been shown to include anti-cancer, anti-angiogenic, and anti-metastatic effects, as well as being able to provide hepatic and neuroprotective effects, immunomodulation, vasodilation, promotion of insulin secretion, and antioxidant activity. Therefore, the purpose of this study was to examine how quickly the ginsenosides decompose and what kinds of degradation products are created under physicochemical processing conditions that don't involve toxic chemicals or other treatments that may be harmful. Methods: The formation of ginsenoside-Rg2 and ginsenoside-Rg3 was examined. These demonstrated diverse pharmacological effects. Results: We also investigated physicochemical factors affecting their conversion. The heating temperatures and times yielding the highest concentration of ginsenosides (-Rb1, -Rb2, -Rc, -Rd, -Rf, -Rg1, and -Re) were examined. Additionally, the heating temperatures and rates of conversion of these ginsenosides into new 'ginseng saponins', were examined. Conclusion: In conclusion, obtained provide us with effective technology to control the concentration of both ginsenosides and the downstream converted saponins (ginsenoside-Rg2, Rg3, Rg5, and Rk1 etc.), as well as identifying the processing conditions which enable an enrichment in concentration of these compounds.

• Journal of Microbiology and Biotechnology
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• 제26권10호
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• pp.1661-1667
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• 2016

The ginsenoside-hydrolyzing β-glucosidase gene (bgy2) was cloned from Lactobacillus brevis. We expressed this gene in Escherichia coli BL21(DE3), isolated the resulting protein, and then utilized the enzyme for the biotransformation of ginsenosides. The bgy2 gene contains 2,223 bp, and encodes a protein of 741 amino acids that is a member of glycosyl hydrolase family 3. β-Glucosidase (Bgy2) cleaved the outer glucose moieties of ginsenosides at the C-20 position, and the inner glucose at the C-3 position. Under optimal conditions (pH 7.0, 30℃), we used 0.1 mg/ml Bgy2 in 20 mM sodium phosphate buffer (PBS) for enzymatic studies. In these conditions, 1.0 mg/ml ginsenoside Rb1 and ginsenoside F2 were converted into 0.59 mg/ml ginsenoside Rd and 0.72mg/ml compound K, with molar conversion productivities of 69% and 91%, respectively. In pharmaceutical and commercial industries, this recombinant Bgy2 would be suitable for producting ginsenoside Rd and compound K.

• Journal of Ginseng Research
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• 제37권1호
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• pp.124-134
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• 2013

The authentication of the physico-chemical properties of ginsenosides reference materials as well as qualitative and quantitative batch analytical data based on validated analytical procedures is a prerequisite for certifying good manufacturing practice (GMP). Ginsenoside Rb1 and Rg1, representing protopanaxadiol and protopanaxatriol ginsenosides, respectively, are accepted as marker substances in quality control standards worldwide. However, the current analytical methods for these two compounds recommended by Korean, Chinese, European, and Japanese pharmacopoeia do not apply to red ginseng preparations, particularly the extract, because of the relatively low content of the two agents in red ginseng compared to white ginseng. In manufacturing fresh ginseng into red ginseng products, ginseng roots are exposed to a high temperature for many hours, and the naturally occurring ginsenoside Rb1 and Rg1 are converted to artifact ginsenosides such as Rg3, Rg5, Rh1, and Rh2 during the heating process. The analysis of ginsenosides in commercially available ginseng products in Korea led us to propose the inclusion of the (20S)- and (20R)-ginsenoside Rg3, including ginsenoside Rb1 and Rg1, as additional reference materials for ginseng preparations. (20S)- and (20R)-ginsenoside Rg3 were isolated by Diaion HP-20 adsorption chromatography, silica gel flash chromatography, recrystallization, and preparative HPLC. HPLC fractions corresponding to those two ginsenosides were recrystallized in appropriate solvents for the analysis of physico-chemical properties. Documentation of those isolated ginsenosides was achieved according to the method proposed by Gaedcke and Steinhoff. The ginsenosides were subjected to analyses of their general characteristics, identification, purity, content quantification, and mass balance tests. The isolated ginsenosides showed 100% purity when determined by the three HPLC systems. Also, the water content was found to be 0.534% for (20S)-Rg3 and 0.920% for (20R)-Rg3, meaning that the net mass balances for (20S)-Rg3 and (20R)-Rg3 were 99.466% and 99.080%, respectively. From these results, we could assess and propose a full spectrum of physico-chemical properties of (20S)- and (20R)-ginsenoside Rg3 as standard reference materials for GMP-based quality control.

• 한국미생물·생명공학회지
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• 제50권3호
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• pp.395-403
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• 2022

본 연구에서는 한국형 프로바이오틱스와 홍삼을 발효하여 저분자 진세노사이드인 compound K (CK)로 생물전환되는지를 확인하였다. 프로바이오틱스 19종의 유전체 분석결과, 진세노사이드 Rb1에서 CK로 전환에 관련된 β-glucosidase는 19종 모든 균주에서 확인되었고, α-arabinofuranosidase 유전자는 3종의 균주, β-xylosidase는 6종 균주, α-rhamnosidase는 8종의 균주에서 확인되었다. 이 중 B. longum CBT BG7는 Rb1으로부터 CK까지 전환시켜, CK 함량을 증가시켰다. 또한, B. breve CBT BR3와 B. lactis CBT BL3은 Rb1을 Rd로 전환시켰다. 균체를 파쇄 또는 미파쇄하여 진세노사이드 전환 반응을 비교했을 때 미파쇄물이 F2와 CK로의 높은 전환량과 수율을 보였다. CBT BG7 + BL3와 BG7 + BR3 혼합균주는 CBT BG7 단독보다 진세노사이드 F2의 함량을 증가시켰다. CBT BG7과 α-amylase 효소를 함께 반응하였을 때에 F2 함량이 증가되었다. 본 연구는 한국형 프로바이오틱스인 CBT BG7, BR3, BL3와 홍삼을 함께 섭취할 경우, 건강에 도움을 주는 생리활성물질인 CK의 생산을 확인하였다. 추후 부탄올 등 다양한 추출용매를 활용하여 생물전환 효율 및 CK로의 전환율에 대한 추가 연구가 필요해 보인다.