• Journal of Ginseng Research
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• v.39 no.3
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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.

• Korean Journal of Food Science and Technology
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• v.15 no.1
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• pp.66-69
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• 1983

Triglyceride fraction was separated from olive oil by thin layer chromatography (TLC) and fractionated into four groups by high performance liquid chromatography (HPLC). Compositions of the triglycerides and fatty acids of four fractions were determined by gas liquid chromatography (GLC). The olive oil contained higher concentrations of C-52 and C-54 triglycerides having partition numbers of 48. The fatty acid compositions of these triglycerides were mainly composed of C18:1 and C18:2 fatty acids. From these results, the possible fatty acid combinations of major triglycerides of olive oil were estimated to be(3C18:1;50.6%), (1C16:0, 2C18:1;23.51%), (2C18:1, 1C18:2;5.48%), (1C18:0, 2 18:1;4.55%), (1C16:0, 1C18:1, 1C18:2;2.94%), (2C16:0, 1C18:1;2.35%), (1 C16:1, 2 C18:1;2.21%), (1C18:1, 2C18:2;1.06%), (1 C14:0, 2 C18:1;1.03%).

• Korean Journal of Weed Science
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• v.7 no.3
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• pp.289-298
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• 1987

This study was conducted to investigate the presence of biologically active substances such as phenolic acids, fatty acids and organic acids in the medical plants like Kalopanax pictum and Acanthopanacis cortex. Alcohol extracts of K. pictum and A. cortex showed complete inhibition of lettuce seed germination, indicating that these plants contained the biologically active substances. Eleven phenolic acids including protocatechuic acid were identified from K. pictum and A. cortex by GLC, and the contents of total phenolic acid were 1.7917mg/g in K. pictum, and 0.9567mg/g in A. cortex. Polyphenols such as neochlorogenic acid, chi orogenic acid, scopoletin, rutin and kaempferolglycoside which were not detected by GLC were anayzed by HPLC, and among phenolic acids indentified chi orogenic acid seemed to be the major acid in both K. pictum and A. cortex presented in amount of 23.7 and 13.0ppm, respectively. K. pictum contained 5.26mg/g of fatty acids and 27.69mg/g of organic acids, and A. cortex possessed 3.22mg/g of fatty acids and 9.80mg/g of organic acids, linoleic and oxalic acid appeared to be the major fatty and organic acids, representing more than 50% of total fatty acids and 80% of total organic acids.

• Biomedical Science Letters
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• v.9 no.4
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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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• v.22 no.4
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• pp.234-238
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• 2007

We used electrospary ionization ion trap tandem mass spectrometry (ESI-IT tandem MS) to structural elucidation of three different biantennary-type glycans having zero, one, two galactoses (G0, G1, G2). The highest fragment ion in the MS/MS spectra of three glycans was produced by 0,2-ring cleavage of fucose-linked N-acetylglucosamine (GlcNAc) in reducing end. The fragment ions both from precursor ions and 0,2-ring cleaved ions ($^{0.2}An$; n=5 for G0, n=6 for G1 and G2) were not overlapped each other. As results of $MS^n$ analyses, tandem fragmentation trees of each glycans were generated and 2,4-ring cleavages ($^{2.4}A_6$) were occurred in GlcNAc linked to reducing end GlcNAc. This structural elucidation and fragmentation study of N-linked glycans by tandem mass spectrometry can be applied to structural analysis of more complicated glycans.

• Korean Journal of Food Science and Technology
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• v.16 no.2
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• pp.179-181
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• 1984

The present study was directed to define the triglyceride composition of pine nut oil. The triglycerides were separated from pine nut oil by thin layer chromatography, and fractionated by high performance liquid chromatography on the basis of partition numbers. Each of these collected fractions were fractionated again by gas liquid chromatography (GLC) according to the acyl carbon number of the triglyceride, and fatty acid composition of the triglyceride was also analyzed by GLC. The pine nut oil consisted of thirty two kinds of triglycerides, and the major triglycerides of pine nut oil were those of $(C_{18:2},\;C_{18:2},\;C_{18:3}\;;\;34.9%)$, $(C_{18:1},\;C_{18:2},\;C_{18:3}\;;\;10.8%)$, $(C_{18:1},\;C_{18:1},\;C_{18:2}\;;\;9.9%)$, $(C_{18:1},\;C_{18:1},\;C_{18:1}\;;\;6.5%)$, $(C_{18:1},\;C_{18:1},\;C_{18:2}\;;\;6.3%)$, $(C_{18:1},\;C_{18:1},\;C_{18:3}\;;\;4.8%)$, $(C_{16:0},\;C_{18:2},\;C_{18:3}\;;\;3.3%)$, $(C_{18:0},\;C_{18:1},\;C_{18:2}\;;\;2.7%)$, $(C_{16:0},\;C_{18:1},\;C_{18:2}\;;\;2.6%)$, $(C_{16:0},\;C_{18:2},\;C_{18:2}\;;\;2.2%)$, $(C_{16:0},\;C_{18:1},\;C_{18:3}\;;\;1.9%)$, $(C_{16:0},\;C_{18:2},\;C_{18:2}\;;\;1.7%)$, $(C_{16:0},\;C_{18:1},\;C_{18:1}\;;\;1.7%)$, $(C_{18:1},\;C_{18:3},\;C_{18:3}\;;\;1.5%)$.

• Korean Journal of Environmental Agriculture
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• v.23 no.3
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• pp.148-157
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• 2004

A rapid analytical method was developed to determine multiple pesticide residues in apples and pears using gas-liquid chromatography (GLC). The samples were extracted with water-miscible solvents and purified by cleanup procedures serially comprising liquid-liquid partition and solid-phase extraction (SPE). Each analyte was separated and determined by a high-resolution GLC equipped with electron-capture detector (ECD) and nitrogen-phosphorous detector (NPD). A total of 196 pesticides, which were previously classified into 5 groups each for ECD and NPD based on their retention behaviors on the capillary column and responses to the detector, were subjected to the recovery experiment. In compliance with the analytical criteria, 70 to 120% of recovery and less than 20% relative standard deviation the proposed method could be successfully applied to analyze 136 and 133 pesticide residues in apples and pears, respectively, which enabled not only rapid screening but quantitation of the residues. Even though less reliability was resulted from unacceptable recovery range, rest of pesticides including 43 and 45 analyzes in apples and pears, could be also detected for their identity. The proposed method fliled to cover 17 and 18 pesticides for apples and pears, which mostly showed high polarity or heat-lability but could be suitable far fast surveilance or monitoring of fruit harvests.

• Proceedings of the Zoological Society Korea Conference
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• 1999.10b
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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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• v.58 no.1
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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.

• The Korean Journal of Mycology
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• v.7 no.1
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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.