Introduction The ginseng saponin (ginsenoside) is one of the most important secondary metabolites in ginseng and hasvarious pharmacological activities. To date about 38 kinds of ginsenosides have been isolated and identified from Panax ginseng C. A. Meyer. Among these ginsenosides, Rg3 is a precursor for ginsenoside Rh2, which has a very strong antitumor effect. and has many pharmaceutical activities. However, Rg3 is extremely low in normal ginseng. Thus production of ginsenoside Rg3 would be very important and many studies have aimed to convert major ginsenosides to the more active minor ginsenoside Rg3. The enzymatic conversion through sugar hydrolysis at a specific position is desirable for the production of active minor ginsenoside Rg3. Material and Method The isolation of β-glucosidase-producing microorganisms was performed according to a previously published method. Each microbialsuspension cultured in nutrient broth was added to the same volume of 1 mM ginsenoside Rb1 solution and then incubated on a rotary shaker at 30°C for 48 h. The reaction mixture was extracted with butanol saturated with H2O and then analyzed by thin layer chromatography (TLC). 8 μl of the ginseng extract solution was spotted on a TLC plate and developed to 5.5 cm distance in a chamber with chloroform/methanol/water as the mobile phase. Bands on the TLC plates were detected by spraying 10% H2SO4, followed by heating. Result and Discussion Ginseng(the root of Panax ginseng C. A. Meyer, Araliaceae) is frequently used as a crude substance taken orally in Korea, China and Japan, as well as other Asian countries, as a traditional medicine. Ginsenosides are the principal components having pharmacological and biological activities. More than 38 different ginsenosides so far have been isolated and identified from ginseng saponins. Among them, deglycosylated ginsenosides are known to be more effective in vivo physiological action and to act as active compounds. A lactic acid bacteria, which have β-glucosidase activity, were isolated from soil and kimchi using a MRS-Esculin agar. These strains were identified on the basis of phylogenetic inference based on 16S rDNA sequences. TLC and HPLC were used to analysis transformed ginsenosides. Ginsenosides are main pharmacoactive component in ginseng. When ginseng was orally administered, the absorption of ginsenosides from the gastrointestinal tract are extremely low. In order to improve oral bioavailability, transforming major ginsenosides into more active minor ginsenoside is very important. Caulobacter leidyia GP45 and Micro- bacterium esteraromaticum GS514 were isolated from ginseng field for converting major ginsenosides into minor ginsenosides. In the co-culture of strain GP45 and GS514 with ginsenoside Rb1, produced compound K and ginsenoside Rg3 individually. The transformation pathway of ginsenoside Rb1 were confirmed Rb1⟶Rd⟶F2⟶compound K and Rb1⟶Rd⟶Rg3.

Ginseng (Panax ginseng C.A. Meyer) has been used as an important oriental medicine since ancient times. Ginseng roots, one of the most famous and expensive crude drugs, have been commonly used to promote the quality of life. Cultivation of P. ginseng is difficult because of its long cultivation period of more than four years, its sun shading, absence of recurring cultivation and various diseases. Conventional breeding of P. ginseng is also difficult and impractical since the procedure takes more than 50 years. In view of these facts, biotechnological applications have been considered as an alternative approach for ginseng improvement and propagation and the production of raw materials for medicinal use. The representative secondary compound accumulated in roots of ginseng species is ginsenoside, a triterpenoid saponin. Ginsenosides are considered to be the main bioactive compounds derived from the roots and rhizomes of different Panax species (Araliaceae). The enzyme squalene synthase catalyzes the first step from the central isoprenoid pathway towards sterol and triterpenoid biosynthesis. Both phytosterols and triterpenes in plants are synthesized from the product of cyclization of 2,3-oxidosqualene. The step in ginsenoside synthesis involves cyclization of 2,3-oxidosqualene to oleanane and a dammarene-type triterpene skeleton. Enzymes at the later step of ginsenoside biosynthesis are cytochrome P450s and glycosyltransferases. The genes for biochemical pathways involved in saponin biosynthesis are of considerable interest in the area of ginseng biotechnology. We investigated the roles of squalene synthase (PgSS1) protein on the biosynthesis of phytosterols and triterpenoids. Over-expression of the PgSS1 gene in adventitious roots of transgenic P. ginseng resulted in the up-regulation of the downstream genes, such as squalene epoxidase, beta-amyrin synthase and cycloartenol synthase. Transgenic P. ginseng also exhibited a remarkable increase in the production of phytosterol (beta-sitosterol, stigmasterol, campesterol) and triterpene saponins (ginsenosides). The first committed step in ginsenoside synthesis is the cyclization of 2,3-oxidosqualene to dammarenediol II by the oxidosqualene cyclase (dammarenediol synthase). The gene encoding dammarenediol synthase was characterized by our research group. We reported that ectopic expression of dammarenediol synthase gene (DDS) in yeast mutant (erg7) lacking lanosterol synthase resulted in the production of dammarenediol and hydroxydammarenone which were confirmed by LC/APCIMS. RNA interference (RNAi) of DDS in transgenic P. ginseng resulted in silencing of DDS expression which leads reduction of ginsenosides production to 84.5% in roots. Now we are focused on the characterization of cytochrome P450 for biosynthesis of protopanaxadiol and protopanaxatriol used as backbones for ginsenoside

To develop a clinically available saponin- or sapogenin complex from Oriental medicines, the EtOH extract (KPRG-A) was obtained by extracting from the four crude drugs, Kalopanacis Cortex, Platycodi Radix, Rubi Fructus and Glycyrrhizae Radis. The BuOH fraction (KPRG-B), a crude saponin complex, was prepared by fractionating KPRG-A, which were further completely hydrolyzed to afford the sapogenin complex (KPRG-D). In an attempt to find the antinoicpetive effects of the saponin complex and sapogenin complex, KPRG-C, and -D, were assayed by writhing-, hot plate-, and tail-flick tests using mice or rats. The three samples were also subjected to antiiflammatory tests using serotonin-induced and carrageenan-induced hind paw edema mice and rats, respectively. The three samples significantly reduced inflammations and pains of the experimental animal. The potency were found in the order of KPRG-D> KPRG-C> KPRG-B. The most active sample, KPRG-D, caused no death, no body increase or no anatomical pathlogic change even at 2,000 mg/kg dose. These results suggest that a sapogenin complex, KPRG-D, which was found to contain mainly hederagenin, platycodigenin, polygalacic acid, 23-hydroxytormentic acid, glycyrrhetic acid together with minor triterpene acids, could be a potential candidate for antiinflammatory therapeutics.

Saponin and ginsenosides are very important as medicinal components of ginseng. These components are significantly different according to Panax species, cultivation area, or manufactured products. In this research, we comparatively analyzed the characteri

To investigate the effect of culture conditions on growth and ginsenosides accumulation, we cultured the ginseng hairy root under three different media, white light or ultra-violet irradiation. The MS/B5 medium containning MS basal salt and B5 vitamin was good for the growth and ginsenoside accumulation. The light during the culture period of ginseng hairy root was irradiated. The growth was abundant in the ginseng hairy root cultured in dark. But the ginsenosides accumulation was higher than in the ginseng hairy root cultured in the light irradiation. When the ginseng hairy root was cultured in 20 L bioreactor, the ginsenosides accumulation was observed at 34% higher than the hairy root cultured in dark. UV irradiated the ginseng hairy root during the culture period. The long time irradiation of UV was caused decreasing the growth of ginseng hairy root, but the accumulation of ginsenosidess was increased as to the irradiated time.

고기능성 홍삼사포닌성분의 함량을 증대시키기 위한 목적으로 홍삼엑스에 열처리, 산(acid)처리하여 그 가능성을 조사하였다. 산도를 조정하지 않은 무처리구(control, pH 4.4)에 120℃ 열처리한 경우 ginsenoside-Rg3의 함량이 약 2배 정도 증가였다. 구연산으로 pH 2.0으로 조정하고 온도처리한 처리구에서는 2.8배나 많은 ginsenoside-Rg3 성분이 증가하였으나 다른 유효한 사포닌의 파괴가 두드러져 처음 홍삼엑스에 함유되어 있던 총사포닌의 65% 정도가 소실되었다. 80℃에서 12시간 처리를 한 경우에는 pH를 2.5와 2.0로 조정한 처리구에서는 11.20 mg과 12.50 mg으로 홍삼엑스의 3.3 mg보다 3.3배 이상 ginsenoside-Rg3 성분이 변환되었다. Ginsenoside-Rb1, Rb2, Rc, Re, Rg1의 함량이 산도가 높아짐에 따라서 급격히 소실되었고 홍삼 특이성분(ginsenoside-Rg3, RH2, Rh1)의 함량은 현저히 증가되었다. 매실엑스로 pH를 2.5로 조정한 처리구에서는 13.34 mg으로 홍삼엑스의 3.3 mg보다 4배 이상 변환된 것으로 분석되었다. 비록 31%정도의 total saponin의 감소가 있었으나 120℃의 고온처리에서 처럼 다른 유효한 사포닌의 큰 손실 없이 60℃에 12시간 처리하는 것만으로도 다량의 ginsenoside-Rg3를 생산하는 것을 확인하였다.

홍삼 제조공정을 단순화하면서 효율적으로 유효성분을 증대시키기 위해서 4년근 수삼을 구입하여 깨끗이 수세한 후 96-98℃에 3시간 정도 수증기로 증삼한 후 30시간 정도 열풍건조하여 홍삼제조를 위한 간이법을 개발하였다. 제조한 홍삼으로부터 홍삼엑스(60˚ brix 이상)를 제조하였으며 홍삼엑스의 추출수율은 60% 이상이었다. 간이법으로 제조한 홍삼을 재료로 하여 농축한 홍삼엑스의 사포닌 함량을 조사하기 위해서 HPLC를 이용하여 총 사포닌(total saponin)을 분석하였다. 그 결과 PD계 사포닌 중 암세포전이억제효과와 평활근이완작용이 탁월한 ginsenoside-Rg3에는 H사에 비하여 2배 이상 검출되었으며, 간상해 억제작용과 혈소판 응집억제작용이 있는 것으로 알려진 ginsenoside-Rh1종의 경우 3배 이상 검출이 되었다.

금산 및 음성지역의 4년생 인삼재배 농가포장에서 직파재배 5개소와 이식재배 5개소를 임의로 선정하여 직파와 이식재배에 따른 생육특성 및 엑스와 조사포닌 함량을 비교한 결과는 다음과 같다. 직파재배는 4년근 생존율이 평균 48%로 이식재배의 86%보다 떨어지나 입모수가 평균 96주/3.3m2순로 이식재배의 57주보다 많고 엽면적지수가 커 수량성이 높은 반면, 주당근중은 작았다. 직파재배는 이식재배에 비해 동체의 신장이 양호하나 지근의 발달이 불량하여 동체중의 비율이 높고 지근중의 비율이 낮았으며, 직파재배는 적변 발생율이 적으나 동체와 지근부위의 엑스와 조사포닌 함량이 낮았다.

인삼원료삼의 안정적인 공급을 위하여 인삼의 기내배양에 의하여 생산된 캘루스 및 모상근으로부터 인삼사포닌, 지방산, 산상다당체, 페놀성화합물 및 유기산함량을 조사하였다. 인삼사포닌의 경우에는 캘루스에 비해 모상근에서 훨씬 많았으며 모상근 세포주간에는 약 10mg/g로 유사한 경향을 보였다. 포화지방산은 인삼 캘루스에서 높은 반면 모상근의 경우에는 불포화지방산이 오히려 많거나 비슷한 경향을 보였다. 특히 인삼 캘루스에서는 palmitic acid가 많이 존재하였으며, 인삼 모상근의 경우에는 불포화지방산인 linoleic acid가 캘루스의 5배 이상 많은 검출되었다. 산성 다당체와 페놀성화합물은 기내 배양한 인삼 캘루스 조직에서는 거의 동량으로 존재하였으나, 인삼 모상근의 경우에는 세포주에 따라서 많은 차이를 나타내었다. 특히 HR-2같은 경우에는 산성 다당체보다는 페놀화합물이 2.26%로 산성 다당체보다 2배 이상 많이 존재하였는데 반하여, HR-3 인삼 모상근의 경우에는 산성 다당체가 1.64%로 페놀화합물보다 4배 이상 존재하는 것으로 분석되었다. 인삼 모상근세포주와 캘루스의 유기산은 모상근 세포주가 34.64 mg으로 가장 많이 존재하는 것으로 조사되었다.

Plants have been known to accumulate a very diverse range of triterpene saponins. We have investigated the regulation of saponin biosynthesis in higher plants using Centella asiatica (L.) Urban as a model plant. Effects of a feeding precursor on asiaticoside production from leaves and on the level of two-type OSCs mRNA were investigated. As a feeding precursor, squalene negatively affected the levels of CYS and bAS mRNA, but it also decreased the production of asiaticoside from whole plants. Plant hormones regulate secondary metabolism, and in plant tissue cultures they could affect both culture growth and secondary metabolite production. Although enhancement of asiaticoside production from whole plant cultures by addition of TDZ (thidiazuron) has been reported, the positive effect of TDZ on the levels of OSCs transcripts was not observed.

There is much evidence suggesting that compounds present in soybean can prevent cancer in many different organ systems. Especially, soybean is one of the most important source of dietary saponins, which have been considered as possible anticarcinogens to inhibit tumor development and major active components contributing to the cholesterol-towering effect. Also they were reported to inhibit of the infectivity of the AIDS virus (HIV) and the Epstein-Barr virus. The biological activity of saponins depend on their specific chemical structures. Various types of triterpenoid saponins are present in soy-bean seeds. Among them, group B soyasaponis were found as the primary soyasaponins present in soybean, and th e 2, 3-dihydro-2, 5-dihydroxy-6- methyl-4H-pyran-4-one(DDMP)-conjugated soyasaponin α~textrmg , β~textrmg , and β a were the genuine group B saponins, which have health benefits. On the other hand, group A saponins are responsible for the undesirable bitter and astringent taste in soybean. The variation of saponin composition in soybean seeds is explained by different combinations of 9 alleles of 4 gene loci that control the utilization of soyasapogenol glycosides as substrates. The mode of inheritance of saponin types is explained by a combination of co-dominant, dominant and recessive acting genes. The funtion of theses genes is variety-specific and organ specific. Therefore distribution of various saponins types was different according to seed tissues. Soyasaponin β~textrmg was detected in both parts whereas α~textrmg and β a was detected only in hypocotyls and cotyledons, respectively. Soyasaponins ~gamma g and ~gamma~textrmg were minor saponin constituents in soybean. In case group A saponins were mostly detected in hypocotyls. Also, the total soyasaponin contents varied among different soy-bean varieties and concentrations in the cultivated soy-beans were 2-fold lower than in the wild soybeans. But the contents of soyasaponin were not so influenced by environmental effects. The composition and concentration of soyasaponins were different among the soy products (soybean flour, soycurd, tempeh, soymilk, etc.) depending on the processing conditions.

1. 종자형성 계통군의 출현은 흑색 PE 피복, 녹색 PE 피복 및 투명 PE 피복이 무피복보다 각각 1일, 2일 및 3일 빨랐으며, 유식물체형성 계통군의 출현은 흑색 PE 피복, 녹색 PE피복 및 투명 PE피복이 무피복보다 각각 2일, 2일 및 3일 빨랐다. 2. 생근경중은 종자형성계통 군에서 투명 PE 피복이 무피복보다 16.7% 증수하였으며, 유식물체형성 계통 군에서는 녹색 PE 피복 및 투명 PE 피복이 무피복보다 각각29.4%, 26.5% 증수하였다. 3. 메탄올엑스함량은 종자형성 계통 군에서는 투명 PE피복이 42.60%로서 가장 많았으며, 흑색 PE 피복이37.50%로서 무피복보다 많았으나, 유식물체형성 계통군에서는 각처리 모두 무피복과 차이가 없었다. 4.조사포닌 함량은 종자형성 계통군에서는 투명 PE피복이 8.25%로서 무피복, 흑백 PE 피복 및 녹색 PE피복보다는 높았으나, 흑색 PE 피복과는 차이가 없었으며, 유식물체형성 계통군에서는 투명 PE피복이 7.75%로서 흑색 PE 피복이나, 녹색 PE 피복보다 높은 함량을 나타냈으나, 무피복 및 흑백 PE 피복과는 차이가 없었다.

창포의 뿌리를 주근(main root)과 측근(lateral root)으로 분리하여 3 열풍건조분말의 주된 포화지방산으로는 caprylic acid, pentadecanoic acid, stearic acid 및 heneicosanoic acid 등 10종이 확인되었고, 이들은 각각 주근과 측근의 경우 caprylic acid가 28.35%, 31.44%이었고, 불포화지방산으로는 linoleic acid, palmitoleic acid 및 lin