• Journal of Ginseng Research
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• 제39권3호
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• pp.221-229
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• 2015

Background: Minor ginsenosides, those having low content in ginseng, have higher pharmacological activities. To obtain minor ginsenosides, the biotransformation of American ginseng protopanaxadiol (PPD)-ginsenoside was studied using special ginsenosidase type-I from Aspergillus niger g.848. Methods: DEAE (diethylaminoethyl)-cellulose and polyacrylamide gel electrophoresis were used in enzyme purification, thin-layer chromatography and high performance liquid chromatography (HPLC) were used in enzyme hydrolysis and kinetics; crude enzyme was used in minor ginsenoside preparation from PPD-ginsenoside; the products were separated with silica-gel-column, and recognized by HPLC and NMR (Nuclear Magnetic Resonance). Results: The enzyme molecular weight was 75 kDa; the enzyme firstly hydrolyzed the C-20 position 20-O-${\beta}$-D-Glc of ginsenoside Rb1, then the C-3 position 3-O-${\beta}$-D-Glc with the pathway $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}C-K$. However, the enzyme firstly hydrolyzed C-3 position 3-O-${\beta}$-D-Glc of ginsenoside Rb2 and Rc, finally hydrolyzed 20-O-L-Ara with the pathway $Rb2{\rightarrow}C-O{\rightarrow}C-Y{\rightarrow}C-K$, and $Rc{\rightarrow}C-Mc1{\rightarrow}C-Mc{\rightarrow}C-K$. According to enzyme kinetics, $K_m$ and $V_{max}$ of Michaelis-Menten equation, the enzyme reaction velocities on ginsenosides were Rb1 > Rb2 > Rc > Rd. However, the pure enzyme yield was only 3.1%, so crude enzyme was used for minor ginsenoside preparation. When the crude enzyme was reacted in 3% American ginseng PPD-ginsenoside (containing Rb1, Rb2, Rc, and Rd) at $45^{\circ}C$ and pH 5.0 for 18 h, the main products were minor ginsenosides C-Mc, C-Y, F2, and C-K; average molar yields were 43.7% for C-Mc from Rc, 42.4% for C-Y from Rb2, and 69.5% for F2 and C-K from Rb1 and Rd. Conclusion: Four monomer minor ginsenosides were successfully produced (at low-cost) from the PPD-ginsenosides using crude enzyme.

• 한국식품과학회지
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• 제15권1호
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• pp.66-69
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• 1983

올리브기름의 트리-글리세리드조성(組成)을 구명(究明)하기 위하여, 시료유(試料油)를 TLC에 의하여, 트리-글리세리드를 분리(分離)하고, 분리(分離)한 트리-글리세리드 HPLC에 의하여 PN별(別)로 4군(群)으로 분획(分劃)하였으며, 각 획분(劃分)을 분취(分取)하여 GLC에 의하여 acyl탄소수별(炭素數別)로 분획(分劃)하였다. 또 PN별(別) 획분(劃分)은 GLC로 지방산조성(脂肪酸組成)을 분석(分析)하였다. 올리브기름에 있어서 주(主)로 PN48에 acyl탄소(炭素) C-52 및 C-54가 높은 비율(比率)로 함유되어 있으며, 지방산(脂肪酸)은 C18 : 1 및 C18 : 2가 주요성분(主要成分)으로 구성(構成)되여 있었다. 시료유(試料油)의 주요(主要) 트리-글리세리드를 들면 다음과 같다. $(3{\times}C18:1;50.46%),\;(1{\times}C16:0,\;2{\times}C18:1;23.51%)$ $(2{\times}C18:1,\;1{\times}C18:2;5.48%),\;(1{\times}C18:0,\;2{\times}C18:1;4.55%)$ $(1{\times}C16:0,\;1{\times}C18:1,\;1{\times}C18:2;2.94%)\;(2{\times}C16:0,\;1{\times}C18:1;2.35%)$ $(1{\times}C16:1,\;2{\times}C18:1;2.21%),\;(1{\times}C18:1,\;2{\times}C18:2;1.06%)$, 및 $(1{\times}C14:0,\;2{\times}C18:1;1.03%)$이였다.

• 한국잡초학회지
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• 제7권3호
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• pp.289-298
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• 1987

약용식물(藥用植物)을 대상(對象)으로 생리활성(生理活性)을 지닌 phenolic산(酸)과 지방산(脂肪酸) 및 유기산(有機酸) 등(等)을 조사(調査)한 결과(結果)는 다음과 같다. 음나무와 오가피의 EtOH 추출물(抽出物)은 상치의 발아(發芽)를 강하게 억제(抑制)하는 생리활성물질(生理活性物質)을 함유(含有)하고 있었다. 음나무와 오가피로부터 GLC에 의해 동정(同定)된 phenolic 산(酸)은 protocatechuic 산(酸) 외(外) 11종(種)이었으며 총(總) phenolic 산(酸)의 함량(含量)은 음나무가 1.7971mg/g, 오가피가 0.9567mg/g 이었다. HPLC로 검정(檢定)된 polyphenols은 ncochlorogenic, chlorogenic, scopoletin, rutin, kaempferolglycoside 였으며 chlorogenic 산(酸)의 함량(含量)이 음나무 23.7ppm, 오가피 13.0ppm으로 가장 많았다. 음나무는 지방산(脂肪酸)이 5.26mg/g, 유기산(有機酸)이 27.69mg/g, 오가피는 지방산(脂肪酸)이 3.22mg/g, 유기산(有機酸)이 9.80mg/g 함유(含有)되어 있었으며 음나무가 오가피보다 많은 지방산(脂肪酸)과 유기산(有機酸)을 함유(含有)하고 있었다. 지방산(脂肪酸) 가운데는 linoleic 산(酸)이 전체(全體)의 50% 이상(以上)을, 유기산(有機酸) 가운데는 oxalic 산(酸)이 80% 이상(以上)의 비율(比率)로 함유(含有)되어 있었다.

• 대한의생명과학회지
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• 제9권4호
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• pp.177-181
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• 2003

The baculovirus/Spodoptera frugiperda (Sf) cell system has become popular for the production of large amounts of the human erythrocyte glucose transporter, GLUT1, heterologously. However, it was not possible to show that the expressed transporter in insect cells could actually transport glucose. The possible reason for this was that the activity of the endogenous insect glucose transporter was extremely high and so rendered transport activity resulting from the expression of exogenous transporter very difficult to detect. Sf21-AE cells are commonly employed as the host permissive cell line to support the baculovirus AcNPV replication and protein synthesis. The cells grow well on TC-100 medium that contains 0.1 % D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, unlike the human glucose transporter, very little is known about properties of the endogenous sugar transporter(s) in insect cells. Thus, the uptake of 2-deoxy-D-glucose (2dGlc) by Sf21-AE cells and the inhibition of 2dGlc transport in the insect cells by fructose and cytochalasin B were investigated in the present work. The binding assay of cytochalasin B was also performed, which could be used as a functional assay for the endogenous glucose transporter(s) in the insect cells. Sf21-AE cells were infected with the recombinant virus AcNPV-GT or no virus, at a multiplicity of infection (MOI) of 5. Infected cells were resuspended in PBS plus and minus 300 mM fructose, and plus and minus 20 $\mu$M cytochalasin B for use in transport assays. Uptake was measured at 28$^{\circ}C$ for 1 min, with final concentration of 1 mM deoxy-D-glucose, 2-[1,2-$^3$H]- or glucose, L-[l,$^3$H]-, used at a specific radioactivity of 4 Ci/mol. The results obtained demonstrated that the sugar uptake in uninfected cells was stereospecific, and was strongly inhibited by fructose but only poorly inhibitable by cytochalasin B. It is therefore suggested that the Sf21-AE glucose transporter has very low affinity for cytochalasin B, a potent inhibitor of human erythrocyte glucose transporter.

• KSBB Journal
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• 제22권4호
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• pp.234-238
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• 2007

본 연구에서는 전극분무 이온화-이온 포획 질량 분석기를 이용하여 인체 IgG의 N-연결 글라이칸 중 이중촉각 구조를 가지면서 비환원 말단의 갈락토오즈 개수가 0, 1, 2 개인 서로 다른 세 가지 글라이칸의 단일 쪼개짐 (MS/MS) 및 다중 쪼개짐 현상을 관찰하고 이를 구조 분석에 이용하였다. MS/MS 분석에서는 퓨코오즈가 결합된 환원 말단의 N-아세틸 글루코사민의 0,2-고리 쪼개짐으로 파생되는 조각 피크가 가장 높은 세기로 나타나는 것을 관찰할 수 있었고, 전구체 피크와 별개로 연속적인 당 단위체의 쪼개짐이 일어나는 것을 알 수 있었다. 또한 G1 글라이칸의 경우에서만 비환원 말단의 갈락토오즈와 N-아세틸글루코사민이 결합된 채 쪼개지는 현상이 일어나는 것을 관찰할 수 있었다. 다중 쪼개짐 질량 분석 기법을 이용하여 MS/MS 스펙트럼에서 나타나는 조각 피크들의 구조를 재확인할 수 있었고, 이를 트리 구조로 정리할 수 있었다. 또한 추가적인 2,4-고리 쪼개짐 현상이 환원 말단 하나 바깥쪽의 N-아세틸 글루코사민에서 공통적으로 일어나는 것을 관찰할 수 있었다. 이와 같은 다중 쪼개짐 질량 분석기법을 이용하여 보다 복잡한 구조의 글라이칸 구조 분석에 이용될 수 있을 것으로 기대된다.

• 한국식품과학회지
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• 제16권2호
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• pp.179-181
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• 1984

잣기름을 TLC에 의하여 트리 - 글리세리드를 분리하고, HPLC에 의하여 PN별(別)로 트리 - 글러세리드를 분획(分劃)하여 분취(分取)하였다. 분취(分取)한 각획분(各劃分)을 GLC 에 의하여 아실탄소수별(炭素數別)로 재분획(再分劃)하는 한편, 각획분(各劃分)의 지방산조성(脂肪酸組成)을 분석(分析)하였다. 주요(主要) 트리 - 글리세리드를 들면 다음과 같다. $(C_{18:2},\;C_{18:2},\;C_{18:3},\;34.9%)$, $(C_{18:1},\;C_{18:2},\;C_{18:3}\;;\;10.8%)$, $(C_{18:1},\;C_{18:1},\;C_{18:2}\;;\;9.9%)$, $(C_{18:1},\;C_{18:1},\;C_{18:1}\;;\;6.5%)$, $(C_{18:1},\;C_{18:1},\;C_{18:2}\;;\;6.3%)$, $(C_{18:1},\;C_{18:1},\;C_{18:3}\;;\;4.8%)$, $(C_{16:0},\;C_{18:2},\;C_{18:3}\;;\;3.3%)$, $(C_{18:0},\;C_{18:1},\;C_{18:2}\;;\;2.7%)$, $(C_{16:0},\;C_{18:1},\;C_{18:2}\;;\;2.6%)$, $(C_{16:0},\;C_{18:2},\;C_{18:2}\;;\;2.2%)$, $(C_{16:0},\;C_{18:1},\;C_{18:3}\;;\;1.9%)$, $(C_{16:0},\;C_{18:2},\;C_{18:2}\;;\;1.7%)$, $(C_{16:0},\;C_{18:1},\;C_{18:1}\;;\;1.7%)$, $(C_{18:1},\;C_{18:3},\;C_{18:3}\;;\;1.5%)$.

• 한국환경농학회지
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• 제23권3호
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• pp.148-157
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• 2004

사과 및 배 시표에서 농약 다성분의 gas chromatography를 이용한 잔류분석법 확립을 위하여 199가지의 농약을 선정하여 retention time 및 검출기에 따라 ECD 5 그룹 및 NPD 5 그룹의 10개 그룹으로 분류하였다. 시료의 종류에 따른 분석조건을 확립하기 위한 회수율 시험은 농약들의 log $P_{ow}$ 값과 화학적 분류에 따라 총 18개 (ECD 11개, NPD 7개)의 농약을 선정하였다. 예비 실험 후 확립된 분석방법에 따라 10개 그룹의 혼합 표준용액으로 사과 및 배에 대한 회수율시험을 행하였다. 그 결과, 총 196가지의 농약의 70%에 해당하는 사과에서 136개, 배에서 133개의 농약들에서 회수율 70에서 120%의 양호한 결과를 나타내었다. 그러나 사과에서 43개, 배에서 45개의 농약들이 70% 미만의 회수율을 보였고, fenvalerate는 120% 이상의 회수율을 나타내었으며, 사과에서 17개 및 배에서 18개의 농약들은 검출이 되지 않았다. 그러나 확립된 분석법은 SOP에 의한 신속하고 수월한 수행으로 농산물 중의 잔류농약을 검출 및 모니터링하는 목적에 적합하다고 사료된다.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1999년도 한국생물과학협회 학술발표대회
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• pp.11-11
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• 1999

;It has been reported that suspension-cultured rice cells grown on mixed carbon sources of glucose (GIc) and acetate exhibited diauxic growth in which acetate was the preferred carbon source (Lee and Lee, 1996). Carrot (Daucus carota L.) suspension cells, showing a diauxic growth very similar to that of rice cells, were used to delineate the mechanisms underlying this preferential use of acetate over GIc. Uptakes of both GIc and 3-0-methylglucose (3-0MG), a non-metabolizable GIc analogue, were similarly inhibited when acetate or butylate, weak acids which are capable of transporting protons into the cytosol, were present in the uptake assay mixture containing cells harvested during the GIc-utilizing second growth phase. Inhibition of GIc uptake by these weak acids was similar when equivalent experiments were carried out with isolated plasma membranes. It was further shown that Glc uptake, which requires a proper proton gradient across the plasma membranes, was inhibited during the first growth phase by acetate-mediated alkalization of growth medium and/or simultaneous acidification of cytosol. This study strongly suggests that Glc utilization in plant cells is inhibited by co-presenting carbon source(s) which can alter the proton gradient across the plasma membrane. We further examined diauxic growth in culture containing GIc and malate. Unlike the case in the culture with GIc and acetate, carrot cells used GIc first. Malate was utilized only after Glc is depleted from medium. These results indicate that GIc can be a preferred or less-preferred carbon source depending on the competing carbon source. It was noted that malate was not directly taken up by cells. Instead it was converted extracellularly into fumarate which was subsequently transported into cells. During the malate-growth phase malate uptake was negligible, and fumarate uptake was active and pH-sensitive. It was shown that fumarase released into medium was responsible for the extracellular conversion of malate into fumarate. An immunoblot experiments showed that fumarase antibody raised against Arabidopsis fumarase provided positive signals only in medium in malate culture, not in fumarate or GIc cultures. This study demonstrates the first example in that fumarase, a mitochondria marker enzyme, can be present in places other than mitochondria.ndria.

• Journal of Applied Biological Chemistry
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• 제58권1호
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• pp.9-11
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• 2015

Tissue extracts were prepared from liquid-cultured Arabidopsis and reacted with UDP-[$^3H$]-GlcNAc. Phospholipid fractions were then extracted by butanol partitioning. Consecutive thin-layer chromatography identified two glycolipids sensitive to PI-specific phospholipase C, known as early intermediates in glycosylphosphatidylinositol anchor biosynthesis; phosphatidylinositol N-acetylglucosamine and phosphatidylinositol glucosamine.

• 한국균학회지
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• 제7권1호
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• pp.9-11
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• 1979

The carpophores of Lentinus lepideus Fr. which grows wildly in Korea were collected in Gyeong Gi Province to examine the chemical components of the fungus. They were extracted with a mixture of chloroform and methanol. From the extract four spots were detected by TLC. One of these spots was identified as ergosterol by TLC, GLC and chemical tests.