인삼의 접합자배로부터 식물체 재분화 및 Farnesyl Diphosphate Synthase 과발현이 진세노사이드 생합성에 미치는 영향

박홍우 忠南大學校 大學院 2010 국내박사

Ginsenosides and phenolic compounds contents and antioxidant activity of various solvent extracts of processed ginseng [Panax ginseng C. A. Meyer] products

권정웅 건국대학교 대학원 2012 국내석사

We analyzed the contents of total ginsenosides and phenol compounds and as well as the antioxidant activities of water, methanol, ethyl acetate, and butanol extracts of 9 Korean ginseng. These consisted of 3 types of processed ginseng (white, red, and taegeuksam) cultivated in 3 locations (Gangwha, Hongcheon, and Pocheon). White ginseng from Hongcheon had the highest total ginsenoside concentration (5,345.4 g/g), in comparison with those from other regions. In particular, the ginsenosides Ro, Rb1, Rh1, and Rb3 were detected at the highest levels. Of red ginseng and taegeuksam, those from Gangwha had the highest total ginsenoside concentration (6,600.9 g/g and 5,124.3 g/g, respectively). Red ginseng from Gangwha had the highest concentrations of notoginsenoside R1, ginsenoside Rf, Rb1, and Rg1 compared to all other red ginseng. Taegeuksam from Gangwha contained the highest total ginsenoside concentration (5,124.3 g/g). Its concentrations of notoginsenoside R1, ginsenoside Rf, and Rb1 were also higher than those of any other taegeuksam. Among the 3 processed ginseng products, red ginseng contained the highest concentration of total phenolic compounds (923.2 g/g), white ginseng contained the second highest concentration (620.5 g/g), and taegeuksam contained the lowest concentration of phenolic compounds (601.8 g/g). With respect to the cultivation area, processed ginseng products from Pocheon contained the highest concentration of total phenolic compounds (910.7 g/g). The processed ginseng products from Hongcheon had the second highest concentration of total phenolic compounds, and those from Gangwha had the lowest concentration of total phenolic compounds (725.0 g/g and 509.5 g/g, respectively). The antioxidant activity of the processed ginseng from 3 different areas was analyzed using the 1, 1-diphenyl-2- picrylhydrazyl (DPPH) method. Of the water extracts, that of Hongcheon taegeuksam showed the highest DPPH radical-scavenging activity at a concentration of 500 g/g (85.94%). Its methanol extract also showed the highest DPPH radical-scavenging activity (78.80%). Of the ethyl acetate extracts, Pocheon taegeuksam showed the highest DPPH radical-scavenging activity (85.78%). In general, the DPPH radical-scavenging activity was highest in the water extracts followed by ethyl acetate extracts, methanol extracts, and butanol extracts. This study demonstrates that the processing method and cultivation area are important factors affecting the concentrations of total ginsenoside and phenolic compounds as well as antioxidant activity.

Biofunctional characterization of puffed red ginseng

이상준 서울대학교 대학원 2017 국내박사

Ginseng (Panax ginseng C.A. Meyer) has been cultivated and consumed as a medicinal herb in East Asia for a long time. Ginseng has a lot of bioactive components including ginsenosides, polyacetylenes, polysaccharides, and phenolic compounds. Among them, ginsenosides have been regarded as major active components of ginseng and used as index component for the quality control. Many researches have been conducted to develop methods for increasing the pharmacological effect of ginseng by conversion of the dammarane-based saponin by high temperature and high pressure thermal processing. However, it is complicated and time-consuming to extract the active components of ginseng because of its dense texture. Thus, researchers have conducted the studies on the production of expanded ginseng using an extruder and explosive puffing process. This study was designed to examine the effect of puffing process on the biofunctional property of red ginseng. Red ginseng was puffed using a rotary puffing machine at 0.30 MPa. After puffing, the changes in physicochemical properties, antioxidant activity and volatile components in puffed red ginseng were investigated. Puffing process increased the total ginsenoside content including ginsenoside Rg3 with anticancer activity. Extraction yields (16.7-42.2%) from puffed red ginseng were higher than those from non-puffed red ginseng (9.0-32.7%) at all extraction times. When comparing the free sugars and amino acids, the contents of maltose and arginine drastically decreased because puffing process accelerated the reaction of maltose and arginine to produce maltulosyl arginine. Effects of explosive puffing on the changes of volatiles in red ginseng were investigated using headspace-solid phase microextraction (HS-SPME)-gas chromatograph (GC) with a mass selective detector (MS). Formation of porous structures and smaller pieces were clearly observed on the surface of puffed red ginseng by scanning electron microscopy. Total volatiles in puffed red ginseng increased by 87% compared with those in red ginseng. Hexanal, Δ-selinene, and β-panasinsene were major volatiles in red ginseng, whereas α-gurjunene, β-panasinsene, and calarene were main volatiles in puffed red ginseng. Puffing process decreased volatiles from lipid oxidation including aldehydes, ketones, and 2-pentylfuran and increased terpenoids in red ginseng. Selective ion monitoring (SIM) mode for GC/MS results showed that 2-furanmethanol and maltol were present at the concentrations of 0.20 and 0.24%, respectively, in red ginseng and 5.86 and 3.99%, respectively, in puffed red ginseng. Explosive puffing process increased 2-furanmethanol and maltol in puffed red ginseng significantly (p<0.05) with the changes of microstructure. The antioxidant properties of extracts of red ginseng and puffed red ginseng were determined in bulk oil and oil-in-water (O/W) emulsions. Bulk oils were heated at 60°C and 100°C and O/W emulsions were treated under riboflavin photosensitization. In vitro antioxidant assays, including 2,2-diphenyl-1-picrylhudrazyl (DPPH), 2,2’-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS), ferric reducing antioxidant power (FRAP), total phenolic content (TPC), and total flavonoid content (TFC), were also performed. The total ginsenoside contents of extract from red ginseng and puffed red ginseng were 42.33 and 49.22 mg/g, respectively. All results from these in vitro antioxidant assays revealed that extracts of puffed red ginseng had significantly higher antioxidant capacities than those of red ginseng (p<0.05). Generally, extracts of puffed red and red ginseng had antioxidant properties in riboflavin photosensitized O/W emulsions. However, in bulk oil systems, extracts of puffed red and red ginseng inhibited or accelerated rate of lipid oxidation, depending on the treatment temperature and the type of assay used. These results suggest that the puffing process can provide us with an alternative means to produce functional red ginseng products with the additional advantage of reduced processing time. Keywords: puffed red ginseng, volatile component, ginsenoside, antioxidant property, bulk oil, oil-in-water emulsion, radical scavenging activity. Student Number: 2000-30738

초단파 처리에 의한 인삼 지상부의 인삼 사포닌 성분 변화

김신정 숙명여자대학교 원격대학원 2011 국내석사

웰빙에 대한 소비자의 선호도가 높아짐에 따라 자연주의 화장품과 한방 화장품이 지속적으로 강세를 보이면서 천연물 기능성 화장품 원료 개발이 활발히 이루어지고 있다. 인삼은 우리나라의 대표적인 특산물로서, 동양에서 가장 오래된 본초서인 신농본초경에 수재된 이래 2,000여 년 동안 한방약으로 사용되어져 왔으며, 인삼 사포닌(진세노사이드)이 주요 약효 성분으로 알려져 있다. 홍삼을 찔 때 약 0.03%로 소량 생성되는 특유 사포닌 성분인 진세노사이드 Rg2는 자외선에 의한 세포 고사를 DNA 회복 과정을 통해 방지시켜서 주름 개선 효과를 갖는다고 보고되었고, 주름 개선 기능 화장품으로 출시되고 있다. 진세노사이드 Rg2는 인삼 사포닌 배당체(glycoside)의 당이 일부분 열이나, 산에 의해서 가수 분해되어 유리된 형태의 프로사포게닌으로 세포 친화성이 좋고, 세포 흡수가 잘 되어 기능성이 강화된 활성 사포닌으로 차세대 기능성 제품의 신소재로서 기대를 모으고 있다. 본 연구에서는 인삼 프로사포게닌 성분 고농도 함유 제제 개발을 목적으로 인삼 재배의 부산물로서 저가 구입이 가능한 인삼 지상부(열매, 화뢰: 꽃봉오리, 줄기와 잎)에 초단파와 2배 식초로 처리를 했을 때 인삼 사포닌 배당체가 활성 사포닌인 인삼 프로사포게닌 성분으로 전환되는 것을 HPLC법으로 확인하여 진세노사이드 Rg2, -Rg3, -Rg5, -Rg6, -Rh1, -F4 등의 인삼 프로사포게닌 성분을 고농도로 함유하는 제제 개발의 최적 조건을 확인하여 다음과 같은 결과를 얻었다. 첫째, 인삼 열매 제제(MGF)에 20분 초단파 처리 시 진세노사이드 Rg2가 2.2749%로 홍삼의 약 84배의 함유량을 확인했으며, 또한, 항 알레르기 작용이 보고된 진세노사이드 Rh1이 1.2803%의 고농도로 함유되는 제제 개발의 조건을 확인할 수 있었다. 항암 작용, 혈관 확장 작용 등이 보고된 진세노사이드 Rg3는 1분 초단파 처리 시 1.1249%로 홍삼의 56배의 함유량을 확인할 수 있었다. 둘째, 인삼 화뢰 제제(MGFB)에서는 1분 초단파 처리 시 진세노사이드 Rg2가 1.5846%로 홍삼의 약 59배, 9분 초단파 처리 시 진세노사이드 Rg3가 1,1766%로 홍삼의 약 59배의 고농도 함유를 보여주었다. 셋째, 인삼 잎과 줄기 제제(MGSL)는 20분 초단파 처리 시 진세노사이드 Rg3가 0.9055% 함유되는 것을 확인할 수 있었다. 이와 같은 연구결과로부터 인삼열매, 화뢰 및 잎 등 인삼 지상부에 초단파 2배 식초 처리를 처리하여 짧은 시간에 인삼 프로사포게닌 성분(진세노사이드 Rg2, -Rg3, -Rg5, -Rg6, -Rh1, -F4)을 높은 농도로 함유하는 제제 개발이 가능하였다. 본 연구 결과로 얻어진 인삼 프로사포게닌 성분 고농도 함유 제제는 기능성 화장품 및 식품의 기능성 강화 신소재로서 활용이 가능할 것으로 사료되며, 향후, 본 제제를 이용한 새로운 기능성 제품의 개발이 기대되고 있으며, 인삼 재배 농민의 소득 증대와 우리 인삼 산업의 친환경 녹색 성장에도 크게 기여할 것으로 사료된다. The purpose of this study was to develop a new preparation process of ginseng extract with the high concentration of ginsenoside Rg2, the red ginseng special component. Chemical transformation from the ginseng saponin glycosides to the prosapogenin was analyzed by the HPLC. The extracts of aerial part ginseng were processed at the several treatment conditions of the microwave and twice brewing vinegar(about 14% acidity). This result showed that the quantity of ginsenoside Rg2 increased over 2% at the 20 minutes of the pH 2~4 vinegar and microwave ginseng berry ethanol extract than other process time. The result of MGB-20 was the ginseng berry extracts that had been processed with microwave and twice brewing vinegar(about 14% acidity) for 20 minutes gained the highest amount of ginsenoside Rg2(2.2749%), Rh1(1.2803%). The MGB-1 was the highest amount of ginsenoside Rg3(1.1249%) in the ginseng berry extract processed with microwave and twice brewing vinegar(about 14% acidity) for 1 minutes. And also, the result of MGFB-1 acquired the highest amount of ginsenoside Rg2(1.5846%) in the ginseng flower bud extract processed with microwave and twice brewing vinegar for 1 minutes. The result of MGFB-9 acquired the highest amount of ginsenoside Rg3(1,1766%) in the ginseng flower bud extract processed with microwave and twice brewing vinegar for 9 minutes. In contrast, the ginseng stem and leaf extracts that processed with microwave and twice brewing vinegar for 20 minutes gained ginsenoside Rg3 0.9055 percent. And the other kinds of ginseng prosapogenin did not show a higher content. This result indicated that the aerial part of ginseng treated with microwave had the quantity of the ginsenoside Rg2 over 84 times when compared to the amount of the ginsenoside Rg2(was not found in raw and white ginseng) in the generally commercial red ginseng.

Taxonomic Study of Novel Bacteria Isolated from Ginseng Rhizosphere and Heavy Metal Resistance Mediated by a Siderophore Producing Bacterium in Panax ginseng Meyer

HUO, YUE 경희대학교 대학원 2019 국내박사

Panax ginseng is one of the most important medicinal plants in Asian countries and is usually harvested after 5 to 6 years of cultivation in Korea. Heavy metal exposure is a type of abiotic stress that can induce oxidative stress and decrease the quality of the ginseng crop. Utilization of plant growth promoting rhizobacteria (PGPR) can be the one way to induce heavy metal resistance in plants. Siderophore-producing rhizobacteria (SPR) is an important PGPRs to improve nutrient uptake and promote plant growth against heave metal stress. Therefore, SPR may be capable of bioremediating heavy metal contamination. In this study, we aimed to isolate and identify potential SPR on heavy metal resistance in P. ginseng. Based on the culture dependent method and polyphasic approach, the five novel strains isolated from ginseng rhizosphere were belonging to the genus Rhodanobacter, Paraburkholderia, Lysobacter, Ornithinimicrobium and Mesorhizobium, designated as DCY112T, DCY115T, DCY117T, DCY118T, and DCY119T. The screening of five isolates had been carried out by in vitro studies for assessment the potentials of bioconversion of ginsenosides, indole acetic acid (IAA) production, siderophores production, phosphates solubilization, antagonistic activity, and heavy metal resistant activities. Compared with all the tested isolates, the siderophore-producing bacteria Mesorhizobium panacihumi DCY119T had higher heavy metal resistance, especially strong resistance to iron (MICFe: 32mM), therefore, this bacterium was chosen as the SPR candidate for further pot test. To investigate the response of P. ginseng seedlings inoculated with the SPR candidate against Fe toxicity, two experiments was designated as in planta pot tests and in vitro medium tests. In planta- grown 2-year-old ginseng seedlings was inoculated with the SPR candidate, and Fe was added or not to investigate the ability of the SPR to reduce oxidative stress and enhance resistance to heavy metal stress. According to the results, ten minutes dipping of 108 CFU/mL DCY119T was enough to prime 2-years old ginseng seedlings against given Fe stress and increased the biomass and sugars contents to promote ginseng growth. Seedlings treated with the SPR DCY119T could increase antioxidant chemicals (total phenolic, total flavonoid) contents and ROS scavenging genes expression to reduce oxidative stress and prevent Fe toxicity. Secondly, the SPR candidate, DCY119T, was cultured in a variety of Fe-supplemented culture media or Fe-contaminated soil extract media and the response of Fe toxicity was assessed by antioxidant activity, IAA, and siderophores production activities. When Fe concentrations in the culture medium was increased, DCY119T grew until 16mM Fe, DPPH scavenging, IAA and siderophores production activities increased, similar results were obtained for Fe-contaminated soil extract media. Together, these results indicate that DCY119T can produce siderophores and scavenge ROS to reduce Fe-induced toxicity, in addition to providing IAA to promote seedling growth, thereby conferring inoculated ginseng with heavy metal resistance. Inoculation of ginseng seedlings with the siderophore-producing rhizobacteria DCY119T decreased the toxic effects of Fe by reducing oxidative stress and promoting growth, indicating that SPR DCY119T can be used for bioremediation of heavy metal contamination. 인삼은 수 천년 전부터 아시아 여러 나라에서 가장 중요한 약용식물 중 하나로 이용되어왔으며, 한국에서는 5~6년 재배 한 후 수확하고 있다. 긴 재배기간으로 인해 토양내 염류가 집적되고, 토양내 미생물의 다양성이 감소되어 병원성 곰팡이에 의한 감염이 4년근 이후 급속히 증가되어 화학적 토양 살균제 등이 암묵적으로 처리되고있다. 염류장애 및 토양 산성화는 인삼의 경작지, 연작지에서 인삼의 생리적 불균형을 초래하여 더욱 중금속에 취약하게 되어 인삼의 품질을 저하시키는 요인이 되고 있다. 산성화된 토양에서 산화된 철이온은 환경스트레스를 더욱 가속화 시킬 수 있는데 철 이온을 조절할 수 있는 siderophore생성능이 우수한 미생물을 활용하여 인삼 뿌리 내 생성된 ROS를 제거하고, 인삼의 중금속 내성을 증진 시킬 수 있는 새로운 환경 친화적인 산화적 스트레스 극복 방안을 본 연구 결과로 제시하고 있다. Siderophore 생성능이 우수한 인삼 근권에 정착된 신규 세균을 분리하였으며, 이들의 인삼 생육 촉진 효과를 증명하였고 신규한 세균에 의해 인삼의 토양내 중금속 스트레스 내성이 증진효과를 검증하였다. 인삼 근권으로부터 분리된 5개의 새로운 균주는 속명(genus) Rhodanobacter, Paraburkholderia, Lysobacter, Ornithinimicrobium 그리고 Mesorhizobium에 속했으며, DCY112T, DCY115T, DCY117T, DCY118T 그리고 DCY119T로 명명하였다. 분류학적 특성을 바탕으로 진세노사이드 생물전환, IAA 생산, Siderophore 생산, phosphates 가용화, 길항 작용 그리고 중금속 내성 작용의 평가를 위해 분리된 신종균주의 특성을 구명하였다. 또한 in vitro 연구 결과, Siderophore 고생성 균주인 Mesorhizobium panacihumi DCY119T는 강한 중금속 내성을 나타내었으며, 특히 Fe (MICFe: 32mM)에 강한 내성을 보여주었다. 그리고 Mesorhizobium panacihumi DCY119T는 인삼을 이용한 생육실험 결과 철이온에 의한 인삼의 산화적 스트레스를 경감시킬 수 있고 인삼의 세근 발달을 통한 양분 흡수률을 높여 인삼의 생육을 더욱 촉진 시킬 수 있는 생물 제재로 사료된다. 인삼내 Fe 독성을 감소시키는 메커니즘을 조사하기 위해 2년생 묘삼에 접종한 Mesorhizobium panacihumi DCY119T 을 이용하여 in planta 화분(pot)실험과 in vitro 실험을 진행하였다. 이 결과, 2년생 묘삼에 108 CFU/mL DCY119T 10분 처리 시, 처리된Fe 스트레스를 완전히 극복하였고, 묘삼의 바이오매스와 당 성분들이 증가됨을 통해 묘삼의 생장을 촉진하는 것을 확인할 수 있었다. DCY119T가 처리된 묘삼은 항산화 성분 (total phenolic, total flavonoid)과 산화 스트레스를 줄이고 Fe 독성을 막는 ROS scavenging에 관여하는 유전자의 발현이 증대 되었다. 실제 인삼이 재배되는 토양에 Fe 염을 고농도로 처리하고 이를 분획하여 DCY119T를 배양하여 실제 토양내에서 균주의 활력과 생물 촉진 활성이 유지되고 있는지 확인한 결과 상당히 높은 농도가 처리된 토양 추출물에서도 본 균주의 활성은 유지됨을 알 수 있었고, 항산화, IAA, siderophores생산 활성도 상대적 정량 측정할 수 있었다. 배양 배지의 Fe농도 증가에서, DCY119T는 16mM 까지 자랐고, DPPH scavenging, IAA, Siderophore 생산 활성이 증가하였으며, Fe가 오염된 흙 추출물이 함유된 배지에서도 유사한 결과가 도출되었다. 그러므로, Siderophore를 생산하는 DCY119T는 Fe 스트레스의 독성을 줄이기 위해 인삼 뿌리내의 ROS를 감소시키고, 식물의 중금속 내성을 유도한다는 것을 알 수 있었다. 본 연구 결과, Siderophore 생성능이 우수한 신규 미생물 Mesorhizobium panacihumi DCY119T 는 향후 인삼의 산화적 스트레스를 줄일 수 있는 생물학적 제제로 이용될 수 있으며, DCY119T 의 ROS scavenging 활성과 인삼의 외부 중금속 스트레스 경감 작용에 대한 기초 연구를 수행하였으나 보다 명확학 상호 작용 기작은 향후 연구를 통해 구명되어져야 할 것이다.

원적외선 조사에 따른 인삼(Panax ginseng C.A. Meyer)의 이화학적 성분변화

朴亨宰 강원대학교 2009 국내석사

인삼은 한국, 중국, 일본을 비롯한 아시아지역에서 다양한 질병을 치료하는 전통 의약으로 사용되었으며, 인삼의 주요 성분으로는 인삼사포닌(ginsenoside)으로 알려져 있으며, 인삼은 항산화 활성, 면역조절, 항피로, 항염증, 항암 등의 효능을 가지고 있다. 본 연구에서는 원적외선 및 열풍 건조와 증숙 처리한 인삼의 페놀성분과 ginsenoside 함량 및 DPPH free radical 소거활성의 변화를 알아보았다. 건조방법에 따른 총 polyphenol과 총 flavonoid 함량은 열풍이나 증숙처리에 비해 원적외선 처리가 높은 함량을 나타냈으며, DPPH free radical 소거활성 또한 원적외선에서 높은 소거활성을 보였으며, 이는 페놀성분의 증가로 항산화 효과가 증가되었을 것으로 사료된다. ginsenoside 함량은 열풍건조 80℃에서 주요 사포닌이 높은 함량을 보였으나, 120℃에서 ginsenoside Rh₁, Rg₂, Rg₃의 함량이 증가하였다. 원적외선 건조에서는 Rf, Rb₃, Rd를 제외한 ginsenoside의 함량이 감소하였으며, 120℃에서 Rg₃가 검출되었다. 증숙처리에서는 Rf를 제외한 ginsenoside의 함량이 증가하였으며, 120℃에서 ginsenoside Rg₃와 Rh2가 검출되었다. 원적외선 조사시간에 따른 총 polyphenol 함량은 원적외선 건조인삼이 백삼에 비해 20배 이상, 홍삼에 비해 1.6배 이상 높은 함량을 나타내었으며, 총 flavonoid 함량은 원적외선 건조인삼이 백삼에 비해 3.4배 이상, 홍삼에 비해 2.4배 이상 높은 함량을 나타내었다. DPPH free radical 소거활성은 원적외선 건조 인삼이 백삼과 홍삼에 비해 각각 17.6배와 2.7배 이상 높은 소거활성을 보였다. 원적외선 건조 인삼의 ginsenoside 함량은 60분과 90분에서 함량이 증가하나 120분 건조에서는 함량이 감소하였으며, 90분 처리에서 ginsenoside Rg₃가 홍삼보다 높은 함량을 나타내었다. 이상의 결과를 종합해 보면 원적외선 건조에서 높은 페놀성분의 함량을 얻을 수 있으며, 증숙처리 후 건조로 원적외선을 이용하면 높은 ginsenoside 함량을 얻을 수 있을 것으로 사료된다. 또한 건조인삼 제조 시 원적외선 98℃에서 90분간 처리하면 가장 높은 페놀성분과 ginsenoside를 얻을 수 있을 것으로 사료된다. 따라서 앞으로 원적외선을 이용하면 경제적인 효율이 좋고, 유효성분이 높은 건조인삼을 제조할 수 있을 것으로 사료된다. Ginseng (P. Ginseng C.A. Meyer) has been used for traditional medicine in Korea, China, Japan and other asian countries. Ginseng saponin (ginsenosides) have been regarded as the principal components responsible for the pharmacological activity of ginseng. It is well known that the ginseng components are effective as antioxidants, immunomodulation, anti-fatigue, anti-inflammatory and anti-tumor. The content of components and thier pharmacological effects are variable by the treatment of heat. This study was conducted to determine antioxidant activity, total polyphenol, total flavonoid and ginsenoside contents from dried ginseng samples by different drying processes. The dried ginseng samples used in the experiment were treated in three different methods, including heat dry (HD), far-infrared irradiation dry (FIRD) and steam treatment. The experiment resulted that the total polyphenol and total flavonoid contents were higher due to FIR treatment than HD and steam treatment. Likewise, free radical scavenging activity was also higher due to FIR treatment. The evaluation of ginsenoside contents due to different treatments showed that HD at 80℃ treatment exhibited to higher contents of major ginsenosides, including Rg₁, Re, Rf, Rb₁, Rb₂, Rd. However, ginsenoside Rg₂, Rh, and Rg₃ were maximized in HD at 120℃. The ginsenoside contents evaluated in this experiment was relatively lower in FIR treatment except for Rf, Rb₃ and Rd, and Rg₃, as compared to 120℃. The evaluation of ginsenoside contents in steam treatment showed that except Rf, all other ginsenosides were increased. The ginsenoside Rg₃ and Rh2 were detected in FIR at 120℃. Raw ginseng roots were treated with FIR in different interval of time(60, 90 and 120min) at 98℃. The result showed that the total polyphenol and total flavonoid contents were higher in FIR ginseng treatments than those of white ginseng and red ginseng. Similarly, the free radical scavenging activity was also higher due to FIR treatments compare to white ginseng and red ginseng. The evaluation of ginsenoside contents due to the effect of time and temperature showed that all the ginsenosides were increased at 60 and 90 minute of treatment in FIR but decreased at 120 minute of expose at 98℃. similarly, the ginsenoside Rg₃ was higher at 90 minute of treatment then that of red ginseng. In conclusion, FIR treatment to ginseng root for drying process was beneficial to increase contain ginsenosides like Rg₃. Thus, the drying process should be capable to apply a manufacturing tool to development new ginseng products. Overall, our results revealed that the FIR treatment is an efficient method to increase the total polyphenol and total flavonoid contents in dried ginseng. moreover, the treatment of dried ginseng at 98℃ for 90 minute can be used as a standard protocol to increase the total polyphenol, total flavonoid, free radical scavenging activity and ginsenoside contents in ginseng.

다중 표적 프로파일 기법을 이용한 인삼 및 홍삼 가공식품 중 유사원료 혼입 판별 연구

최지연 조선대학교 대학원 2017 국내박사

The consumption of medicinal food to maintain good health has increased, resulting to extend the markets for functional foods. Especially, ginseng(Panax ginseng, ginseng radix) and red ginseng; a processed ginseng product in South Korea, are commonly used as foods in general and healthy functional food in particular from thousands of years ago. Due to high market demands, inexpensive morphologically similar materials may be intentionally mixed into ginseng or red ginseng products to derive benefits. It is very important to know whether the ginseng or red ginseng products are adulterated or not. The genetic analysis cannot be applied to processed red ginseng products, as they involve high temperature and pressure processing and genes may be destroyed. Therefore, this study was designed to find out some physicochemical indicators that would not be changed in various processing processes. Several possible substances such as Codonopsis lanceolata, Platycodon grandiflorum, and Pueraria lobata were tested as adulterants and investigated in the study. To approached with multi-taret profiling method in subject samples and report a suitable method, the inorganic components were analyzed by using inductively coupled plasma/mass spectrometry(ICP/MS) and inductively coupled plasma/optical emission spectrometry(ICP/OES), volatile organic components were analyzed by using gas chromatography-mass spectrometry(GC-MS), phytochemical screening via qualitative and quantitative analyses were done using standard reference methods and analysis of nonvolatile organic components were determined using high performance liquid chromatography(HPLC) and LC-mass spectrometry/mass spectrometry(MS/MS). Six macro elements(Na, Mg, Ca, K, Fe, and Al) in individual dried and steamed dried samples were analyzed through ICP-OES and 19 trace elements(Ba, Cr, Cu, Mn, Ni, Rb, Sr, Zn, Ga, Se, Tl, Be, Co, V, Li, Cs, Bi, Pb, Cd) in the samples were analyzed using ICP-MS. No significant difference due to processing was shown between raw samples and steamed samples and it was difficult to discriminate among the samples based on the analyzed contents of inorganic components. Among the statistical analyses, the Linear Discriminant Analysis (LDA) of the inorganic profile enabled discrimination among the individual samples. The raw samples and steamed samples were clearly distinguished from each other on the graph. Thus discrimination among the samples using inorganic elements was possible by LDA statistics. The volatile organic components were extracted using the Solid phase micro extraction(SPME) method, and analyzed using GC-MS. From the results, (E)-2-hexen-1-ol and barbatene were identified in the raw samples of C. lanceolata and P. gradiflorum, but not in the raw samples of the P. ginseng and P. lobata. Furaneol was detected only from the steamed sample of P. gradiflorum. There were difficulties in the analysis of volatile organic components because the reproducibility was poor and the expected quantities of indicators were very small. Therefore, it was judged that applying volatile organic components as a method to identify the similar materials mixed into processed ginseng or red ginseng products would be difficult. In phytochemical screening tests, terpenoids, phytosterols, phenolic compounds, coumarins, flavonoids, and alkaloids were screened and changes in colors were observed. Since high reactions of flavonoid and phenolic components were identified in the methanol extract of P. lobata, it was mixed into red ginseng extract at ratios of 10, 20, 30, 40 and 50% and screening tests were conducted. The intensity of the colors changed according to the mixing ratio, so it was easy to check whether or not the P. lobata were mixed. Analyses were conducted using HPLC, to explore indicators that do not exist in ginseng but are detected from C. lanceolata, P. grandiflorum and P. lobata. In the results, peak 1, which was detected in C. lanceolate and P. grandiflorum in the same time zone, and peak 2, which was detected only in P. lobata but not in other samples, were identified. Expected indicators were verified by measuring their accurate masses through LC-MS/MS analyses, and as components not detected in ginseng, the same indicator of lobetyolin was identified in C. lanceolate, P. grandiflorum and an indicator ononin was identified in P. lobata. Therefore, it was concluded that the HPLC method developed in this study can identify whether or not C. lanceolate, P. grandiflorum, or P. lobata has been mixed into ginseng or red ginseng. A total of 61 general and health functional foods made of ginseng or red ginseng distributed in the market were monitored and there were no C. lanceolate, P. grandiflorum or P. lobata mixed into the foods. When samples of processed products of C. lanceolate and P. grandiflorum, which are similar materials, were analyzed respectively, the common indicator lobetyolin was identified. Also when samples of processed P. lobata products were analyzed, ononin was detected. The method developed from this research can be used not only for ginseng processing food but also for the quality examination of bonnet, balloon flower roots, and arrochards. In nutshell the indicator of similar materials, basic physicochemical components, macro inorganic and trace inorganic elements, and volatile organic components, were examined. Although it was difficult to set them as a method for clear discrimination, the examination was meaningful for the collection of basic data. Through this study, a method to identify whether or not similar materials were mixed into ginseng or red ginseng products using the HPLC analysis method was established. The identification method using HPLC was developed in this study and can be applied for development of methods to identify other adulterated foods.

Aspergillus oryzae와 당분해효소를 이용한 인삼의 생물전환

김보혜 부산대학교 2006 국내석사

Biotransformation of Korean Ginseng by Aspergillus oryzae and Commercial Glucosidase Kim Bo-Hye Department of Microbiology and Immunology, College of Medicine, Pusan National University, Pusan, Korea Abstract When Ginseng is taken orally as a crude drugs, the metabolites of Ginseng saponin(ginsenoside) is absorbed from the intestines. The clinical basis of Ginseng treatment may be partly associated with differences in intestinal microflora of hosts. Protopanaxadiol-type ginsenosides such as Rb1, Rb2, and Rc are metabolized by intestinal bacteria after oral administration to their final derivative 20-O-β-D-flucopyranosyl-20-(S)-protopanaxadiol (compound K, M1, IH901). The in vivo anti-metastatic effect of the ginsenosides was primarily based on their metabolite compound K. It has been assumed that the glycosides cannot be absorbed from the small intestine and cleavage of the β-glycoside linkage will not occur until the compounds reach the microflora in the large intestine. Biological activity depends on the presence or absence of the glycoside residue. The position and nature of the sugar residue may increase the uptake of the compound in the small intestine. The aglycone is likely to have a greater biological effect than the glycoside. Therefore, Ginseng was fermented by Aspergillus oryzae to get metabolites with increased biological activity, higher digestiveness, and modified structure. Then new biotransformed compounds (WG2-2-1, WG2-2-2) were detected and isolated through several chromatographic techniques. And WG2-2-2 was confirmed to biologically active compound K by TLC, HPLC, and mass spectrum. Also biological activity of biotransformed WG2-2-1 and WG2-2-2 were investigated on antibacterial, cytotoxic, and DPPH radical scavenging assay. WG2-2-2 was more active on aboved biological assays. It suggests that WG2-2-1 is intermediate metabolite transforming to final WG2-2-2 (compound K). Also biotransformation of ginseng saponin or ginseng products by pectinex was performed. Then compounds (PG-1, PG-2, PG-3) were biotranformed from ginseng. And those compounds were isolated by several chromatographic techniques. Finally PG-3 was identified as compound K, which was transformed by Aspergillus oryzae in this research. Also its biological activity was observed to be active on DPPH free radical scavenging and proliferation of tumor cells. But minor biotransformed metabolites were under the investigation to obtain a lot of compounds by scale-up fermentation and enzyme reaction to identify those of structure and observe those of biological activity.

Inhibitory effect of long-term and low temperature fermented Ginseng on passive cutaneous anaphylaxis

Hwang, Seonweon Graduate School, Yonsei University 2019 국내박사

Ginseng (the root of Panax ginseng C.A. Meyer) is a well-known Korean traditional medicine in the Far East and has gained popularity in the west in the last decade. Ginseng has been widely reported to possess various biological activities, including anti-oxidant, anti-cancer, and anti-inflammatory activities. Ginseng contains various phytochemicals, such as ginsenosides (saponins), polyacetylenes, and polyphenolic compounds. Among them, the major components are ginsenosides, which are glycosides with steroids or triterpenes as aglycons. These ginsenosides are an important class of physiologically active compounds found in many herbs which possess anti-inflammatory and anti-tumor activities, such as the inhibition of tumor-induced angiogenesis and the prevention of tumor invasion and metastasis. Recently, studies have been actively carried out to develop highly- functional ginseng products by combining new technologies that maximize the efficacy of ginseng. Fermented ginseng, which changes the structure of ginsenosides through fermentation using enzymes and microorganisms, has received much attention. However, the effect of ginseng fermented by Lactobacillus plantarum on allergic responses has not been elucidated. In the present study, for fermentation, ginseng was incubated with L. plantarum at low temperature (4°C–10°C) for a long time (100 days). The amounts and types of ginsenosides in fermented ginseng extract (FPG) were assessed and compared with their corresponding values in the ginseng extract (PG). Any change in the composition of ginsenosides due to fermentation was also investigated. In the fermentation product, Rb1 (94%), Rg1 (61%), and Rc (50%) were decreased. However, the ginsenosides Rh2, Rh3, and Rk2 were newly detected, and the amounts of the ginsenosides Rd (1.7 time), Rh1 (6.5 time), and F1 (35 time) were increased. Anti-allergic effects of FPG in immunoglobulin E (IgE)-mediated murine cell line in vitro and passive cutaneous anaphylaxis in vivo were investigated. FPG showed higher inhibitory effect than PG against allergic responses in vitro and in vivo. The secretion of β-hexosaminidase and interleukin (IL)-4 from IgE-dinitrophenyl (DNP)-stimulated RBH-2H3 mast cells was significantly (p < 0.05) inhibited by FPG treatment in a concentration dependent manner. Further, mitogen-activated protein kinase kinase 4 (MKK4) activation and c-Jun N-terminal kinase (JNK) phosphorylation, which is the subsequent signal of MKK4, were attenuated by FPG treatment. The superior inhibitory effect of FPG compared with that of PG on allergic response was confirmed by IgE-DNP-induced passive cutaneous anaphylaxis in mouse in vivo model. Accordingly, the fermentation of ginseng with L. plantarum possesses anti-allergic effects in vitro and in vivo conditions. Therefore, it is anticipated that fermented ginseng can be used as a natural material for anti-allergic disorders.

Cytohistological study of the leaf structures of Panax species, and molecular and functional characterization of ginseng UGT72AL1 in Arabidopsis

녹 웨이 윙 전남대학교 2017 국내석사

Ginseng, which belongs to Araliaceae family, is the most popular perennial herb worldwide. For decades, ginseng has gained considerable attention on account of its beneficial health effects. Since the 1900s, ginseng has been commercially utilized to produce bioactive products, which to date have been sold in over 35 countries. However, the 4- to 6-year field cultivation of ginseng is a time-consuming and labor-intensive process. Thus, considerable effort has been made to improve ginseng yield and quality via classical and molecular breeding methods, as well as through improvements in cultivation. To shed light on this process, the cytohistology of the leaf structure of two different ginseng species was studied. In addition, functional characterization of a ginseng glycosytransferase gene, PgUGT72AL1, which might be involved in ginseng saponin biosynthesis, specifically the transfer of a glycosyl group to an acceptor, was performed at the molecular and genetic levels. Both Panax ginseng Meyer and Panax quinquefolius are obligate shade-loving plants, the natural habitats of which are the broad-leaved forests of Eastern Asia and North America. Panax species are easily damaged by photoinhibition when they are exposed to high temperatures or receive insufficient shade. Thus, a cytohistological study of the leaf structures of two of the most well-known Panax species was performed to gain a better understanding of the physiological processes that limit photosynthesis. The mesostructure of both P. ginseng and P. quinquefolius frequently has one layer of non-cylindrical palisade cells and three to four layers of spongy parenchyma cells. P. quinquefolius was found to contain similar number of stomata in the abaxial leaf surface but more tightly appressed enlarged grana stacks than P. ginseng; however, the adaxial surface of the epidermis in P. quinquefolius shows cuticle ridges with a pattern similar to that of P. ginseng. The anatomical leaf structure of both P. ginseng and P. quinquefolius shows that they are typical shade-loving sciophytes. Slight differences in chloroplast structure suggest that the two species can be authenticated using transmission electron microscopic images, and light-resistant cultivar breeding can be performed via the control of photosynthetic efficiency. Glycosylation of natural compounds gives rise to a great diversity of secondary metabolites. Glycosylation steps are implicated not only in plant growth and development but also in plant defense responses to various environmental stresses. This glycosylation process is mediated by members of the multigene glycosyltransferase (GT) family, which catalyze the transfer of single or multiple activated sugars to a wide range of substrates, thus influencing their chemical properties and bioactivities. Although their activities have been recognized for a long time and the genes coding for uridine diphosphate (UDP)-dependent glycosyltransferases (UGTs) have been identified in several higher plants, details of the specific functions of GTs remain elusive. Considering that UGTs are key enzymes in the production of numerous different types of ginsenosides in P. ginseng, we selected a UTG gene as a candidate for further functional characterization. Here, we report that the PgUGT72AL1 gene is highly expressed in the upper root, rhizome, and youngest leaf of ginseng. Stress responsiveness against various abiotic stresses, GUS histological expression, and subcellular localization in Arabidopsis were also examined. Exogenous overexpression of PgUGT72AL1 in Arabidopsis resulted in fused organ in the axillary branches, indicating the role of this gene in plant development.