• Journal of the Korean Magnetic Resonance Society
• /
• v.17 no.2
• /
• pp.67-75
• /
• 2013

ATP synthase produces ATP, a major energy source for metabolic processes in organisms, from ADP and inorganic phosphate in cellular membranes. ATP synthase is known as a rotary motor, in which the c-subunit ring functions as a rotor. In this work, we have tried to develop a more general preparation procedure of thermophilic $F_oc$-ring ($TF_oc$-ring) for NMR measurements. The expression of $TF_oF_1$ is easily affected by various experimental conditions such as temperature, shape and size of a flask, a volume of medium, and shaking rate of an incubator. Accordingly, we have tried to optimize the expression conditions of $TF_oF_1$. $TF_oc$-rings were purified from $TF_oF_1$ according to a reported method. We modified purification procedures to improve purity and yield of $TF_oc$. On top of them, we found a new combination of detergents for the purification at anion-exchange column chromatography. To examine the effect of lipid environments on the structure, the $TF_oc$-rings were reconstituted into two kinds of lipid bilayers, namely, saturated and unsaturated lipid ones. Then, we have compared characteristics of the $TF_oc$-ring structures in these membranes with solid-state NMR.

• Journal of Veterinary Science
• /
• v.21 no.4
• /
• pp.63.1-63.9
• /
• 2020

Background: Canine adenovirus type 2 (CAV-2) induces infectious laryngotracheitis in members of the family Canidae, including dogs. To date, no ELISA kits specific for CAV-2 antibody have been commercialized for dogs in Korea. Objectives: We aimed to develop new indirect enzyme-linked immunosorbent assay (I-ELISA) to perform rapid, accurate serological surveys of CAV-2 in dog serum samples. Methods: In total, 165 serum samples were collected from dogs residing in Chungbuk and Gyeongbuk provinces between 2016 and 2018. The Korean CAV-2, named the APQA1701-40P strain, was propagated in Madin-Darby canine kidney cells and purified in an anion-exchange chromatography column for use as an antigen for I-ELISA. The virus-neutralizing antibody titers of CAV-2 in the dog sera were measured by virus neutralization (VN) test. Results: We compared the results obtained between the VN and new I-ELISA tests. The sensitivity, specificity, and accuracy of new I-ELISA were 98.6%, 86.4% and 97.0% compared with VN test, respectively. New I-ELISA was significantly correlated with VN (r = 0.91). Conclusions: These results indicate that new I-ELISA is useful for sero-surveillance of CAV-2 in dog serum.

• The Korean Journal of Mycology
• /
• v.20 no.4
• /
• pp.324-336
• /
• 1992

On the five interspecific protoplast fusants of Ganoderma lucidum and G. applanatum was the antitumor test performed. The fusant F-2 was selected, to examine the cultured mycelia (protein bound polysaccharide) as antitumor components. When a dose of 20 mg/kg/day of each components purifed from F-2 fusant was, i.p., injected into ICR mice, the inhibition ratio of Fr. II against the solid form of sarcoma 180 increased to 1.5 times as compared with that of their parents. When Fr. II was examined for immunopotentiation activity, it increased the amount of the superoxide anion in activated macrophages to 1.2 times and the count of hemolytic plaque forming cells in the spleen to 4.3 times as compared with that of each control group. Its chemical analysis showed 85.2% polysaccharide which consisted of glucose, galactose, mannose, fucose and xylose, and 0.39% protein of 15 amino acids. The content of hexosamine was 0.39% and the molecular weight of Fr. V was $5.6{\times}10^4$ dalton.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.31 no.3
• /
• pp.527-533
• /
• 2002

In the screening of Korean traditional tea sources for the cellular lysosomal enzyme activity of peritoneal macrophage from mice, CT-0, a cold-water extract from Carthamus tinctorius L., showed the highest macro-phage-stimulating activity. CT-1-IIa-2-1, a purified macrophage-stimulation polysaccharide was obtained by a series of purification steps such as anion exchage chromatography with DEAE-Toyopearl 650M, gel permeation chromatography with Sepharose CL-6B, Sephacryl S-200, and HPLC with Superdex G-75. The molecular weight of homogeneous purified polysaccharide was estimated about 68 kDa. CT-1-IIa-2-1 consisted of xylose 27.44%, arabinose 16.14%, mannose 15.92% and glucose 14.47%. To measure acute toxicity, dose of 50, 100, 500, and 1000 mg/kg were intraperitoneally injected to ICR mice. The LD$\_$50/ was about 397 mg/kg.

• The Korea Journal of Herbology
• /
• v.22 no.4
• /
• pp.261-270
• /
• 2007

Objective : The 5-HMF was not index material suitable to do the quality control of Rehmanniae Radix Preparata. In this study, We estimated the changes of oligosaccharide contents in Rehmanniae Radix Preparata using high-performance anion-exchange chromatography with pulsed amperometric detection(HPAEC-PAD). Methods : The analysis of oligosaccharide was conducted by HPAEC-PAD with Carbopac PA1, $250{\times}4mm$, 5um, and Carbopac PA1 guard column. Column temperature was kept at $30^{\circ}C$. Elution was carried out at 1000 ${\mu}l/min$ with 70mM NaOH and the injection volume was $10{\mu}l$. Each component was detected by PAD. Results : Nine constituents were found from merchandising Rehmanniae Radix Preparata(MR), while seven constituents were found in various processed Rehmanniae Radix Preparata. Not all constituents were defined but stachyose and raffinose were found in all cases. And The most common constituents of Rehmanniae radix was stachyose. In the course of processing, most of stachyose and raffinose were decreased. Stachyose was decreased slowly in the course of processing with rice wine(RR), amomi and rice wine(AR), and crataegi and rice wine(CR). However stachyose was decreased rapidly in the course of processing with fresh rehmannia juice(FR). The method with crataegi and rice wine(CR) showed the smallest decrease of stachyose. And processing method with crataegi and rice wine(CR) showed the most abundant amount for stachyose after the nineth processing. Conclusion : The changes of oligosaccharides in the course of processing were a very important direct barometers to do the quality control and set up a standard of Rehmanniae Radix Preparata.

• Microbiology and Biotechnology Letters
• /
• v.26 no.6
• /
• pp.515-522
• /
• 1998

Plant root rotting fungi, Fusarium solani are suppressed their growth by the chitinase which is produced from the antagonistic soil bacteria. The chitinase producable antagonistic bacterium Pseudomonas sp. 3098 was selected as a powerful biocontrol agent of F. solani from ginseng rhizosphere. The antagonistic Pseudomonas sp. 3098 was able to produce a large amount of extracellular chitinase which is key enzyme in the decomposition of fusarial hypal walls. The chitinase was purified from cultural filtrate of Pseudomonas sp. 3098 by the procedure of ammonium sulfate precipitation, anion exchange chromatography, gel filtration on Bio-Gel P-100, and 1st and 2nd hydroxyapatite chromatography. The molecular mass of the purified enzyme was ca. 45 kDa on SDS-FAGE. The optimal pH and temperature for the activity of purified chitinase were 5.0 and 45$^{\circ}C$, respectively. The enzyme was stable in pH range of 5.0 to 9.0 up to 5$0^{\circ}C$ The enzyme was significantly inhibited by metal compounds such as FeCl$_2$, AgNO$_3$ and HgCl$_2$, and was slightly inhibited by p-CMB, iodoacetic acid, urea, 2,4-DNP and EDTA. The enzyme had ability of digestion on colloidal chitin and chitin from shrimp shell, but could not digest chitosan and chitin from crab shell. Km value of the enzyme was 0.11% on colloidal chitin, and the maximum hydrolysis rate of the enzyme was 34% on colloidal chitin.

• Korean Journal of Pharmacognosy
• /
• v.36 no.2 s.141
• /
• pp.129-135
• /
• 2005

Lectins from Allomyrina dichotoma (ADL) and Bombyx mori (BML) were partially purified by physiological saline extraction, ammonium sulfate fractionation, anion exchange column chromatography on DEAE Sephadex A-50 and gel filtration column chromatography on Sephadex G-200. An assay for cytokine expression was carried out by using reverse transcription polymerase chain reaction(RT-PCR). mRNA isolated from PBMC(human peripheral blood mononuclear cells) were stimulated with ADL(O.D.=0.2) and BML(O.D.=0.1) for various times(1,4,8,24,48 and 72 h) and various cytokine mRNA assessed by RT-PCR were shown as follows: The patterns of bands for IL-1 mRNA of BML were very similar with those from ADL and these bands were decreased along the increasing reaction times after showing a strong band at 1 h. However mRNA expressions for IL-2, IL-6, $IFN{\gamma}$ and $TNF{\alpha}$ showed different patterns between ADL and BML. With the effect of ADL, the expression of IL-2 and IL-6 mRNA were continuously detected until 72 h with the strongest band of IL-2 mRNA at 24 h. The strong bands of $IFN{\gamma}$ mRNA were observed from 4 to 8 h but the strongest one of $TNF{\alpha}$ was just observed at 1 h. Meanwhile with BML, the bands for IL-2 and $IFN{\gamma}$ were increased along the increasing reaction times until 72 h. The strongest bands were showed from 4 to 8 h with IL-6 and at 8 h with $TNF[\alpha}$. To verify quantitatively ELISA was used for assay of protein secretions of the cytokine gene with IL-2 and $IFN{\gamma}$ expressed markedly different in RT-PCR. The highest cytokine secretion for IL-2 was demonstrated at 48 h. The production of $IFN{\gamma}$ was markedly increased at 24 h and secreted highest at 72 h. These result suggest that ADL and BML, as inducers of cytokines, can elicit detectable cytokine mRNA from PBMC within the first few hours of stimulation and maintain the production of cytokines for a few days by the methods of RT-PCR and ELISA.

• Food Science and Preservation
• /
• v.16 no.5
• /
• pp.612-617
• /
• 2009

Kimchi is a traditional pickled food using Korean Chinese cabbage(Brassica campestris var. pekinensis) and also containing phytochemicals, glucosinolates. This study was carried out to investigate the changes in the total glucosinolates levels of Kimchi using Korean Chinese cabbage of harvest time(June-July, August-September, October-November, December-April, May) according to storage temperature(4, 10, $15^{\circ}C$) and storage duration(0, 1, 3, 5, 7, 10, 14, 21 and 28 day). For determination of glucosinolates, 50g of Baechu kimchi was used for analytical sample preparation provided with an anion exchanges column and measured by UV-visible Spectrophotometer. The highest contents of water occurred at August-September during fermentation. At 15, 10, $4^{\circ}C$, the pH in all of seasonal variation of Baechu kimchi declined, especially from 0 to 3 days at $15^{\circ}C$, from 0 to 7 days at $10^{\circ}C$ and from 0 to 14 days at $4^{\circ}C$. At that storage, the total glucosinolates levels in all of seasonal variation of Kimchi declined in storage temperature. Baechu kimchi at August-September showed the highest total glucosinolates levels. Also total glucosinolates levels decreased as storage period increased. Baechu kimchi fermented at $15^{\circ}C$ for 7days decreased rapidly and reached to the lowest at the 1day(ranged from $10.3{\pm}0.70$ to $23.4{\pm}0.37{\mu}mol/g$ dry weight). At $10^{\circ}C$ for 14days were ranged from $12.9{\pm}0.29$ to $33.7{\pm}1.81{\mu}mol/g$ dry weight before fermentation and decreased rapidly at the 3day(ranged from $9.5{\pm}0.54$ to $20.5{\pm}0.61{\mu}mol/g$ dry weight). Total glucosinolates levels of Baechu kimchi fermented at $4^{\circ}C$ for 28days decreased rapidly during 7day(ranged from $9.9{\pm}0.78$ to $21.1{\pm}0.96{\mu}mol/g$ dry weight) and then slowly decreased. Therefore the total glucosinolates levels decreased during storage time and depended on harvest time and storage conditions of Kimchi.

• Korean Journal of Environmental Agriculture
• /
• v.16 no.1
• /
• pp.37-43
• /
• 1997

Tolerance mechanism to simazine (6-chloro-N,N'-diethyl-1,3,5-triazine-2,4-diamine) in Coix lacryma-jobi was investigated with respect to herbicide detoxification via glutathione conjugation. Simazine was initially absorbed by seedlings of C. lacryma-jobi and corn, but after 12 hours of treatment, no significant difference in simazine absorption was found in both species. Simazine absorbed was rapidly metabolized to glutathione-simazine conjugate. One to six hours after treatment, metabolism was approximately 2-fold faster in C. lacryma-jobi than in corn. Glutathione content was found 1.5- and 2.3-fold higher in coleoptile and root of C. lacryma-jobi, respectively, compared with corn. In both species, the highest concentration of glutathione was found in coleoptile tissue. Glutathione S-transferase that exhibits activity with 1-chloro-2,4-dinitrobenzene was not significantly different between two species. However, glutathione S-transferase activity with simazine was approximately 2-fold greater in C. lacryma-jobi than in corn. The glutathione S-transferase activity was 20 to 30% greater in shoot of either species than in root. Fast protein liquid chromatography-anion exchange column was used to separate glutathione S-transferase isozymes in coleoptiles of C. lacryma-jobi and corn. A peak of glutathione S-transferase activity with 1-chloro-2,4-dinitrobenzene and two peaks of glutathione S-transferase activity with simazine from C. lacryma-jobi were coeluted with those from corn, but showed greater activity than in the case of corn. Another glutathione S-transferase isozyme that exhibits activity with simazine was detected in the elution of C. lacryma-jobi extract, but not in corn. Electron transport in chloroplast thylakoids isolated from leaves of both species was equally sensitive to simazine applied at 1 to 100 nM. These results indicate that the simazine tolerance in C. lacryma-jobi is due to its capacity to detoxify the herbicide via glutathione conjugation, which is positively correlated with the level of glutathione content and glutathione S-transferase activity.

• Korean Journal of Environmental Agriculture
• /
• v.32 no.4
• /
• pp.315-322
• /
• 2013

BACKGROUND: This study was performed for the identification and quantification of glucosinolate (GSL) contents in seven varieties of rapeseed (Brassica napus L.) sprouts cultivated under dark and light conditions. METHODS AND RESULTS: Crude glucosinolates (GSLs) were desulfated by treating with aryl sulfatase and purified using diethylaminoethyl sepharose (DEAE) anion exchange column. Individual GSLs were quantified using high-performance liquid chromatography (HPLC) with electrospray ionization-tandem mass spectrometry (ESI-MS/MS). Eleven GSLs including six aliphatic (progoitrin, sinigrin, glucoalyssin, gluconapoleiferin, gluconapin, and glucobrassicanapin), four indolyl (4-hydroxyglucobrassicin, glucobrassicin, 4-methoxyglucobrassicin, and neoglucobrassicin) and one aromatic (gluconasturtiin) were identified based on the fragmentation patterns of MS spectrum. Aliphatic GSLs were noted as the predominant group with average 85.2% of the total contents. The most abundant GSLs were progoitrin which was ranged at $8.14-118.68{\mu}mol/g$ dry weight (DW). The highest total GSL amounts were documented in 'Hanra' ($146.02{\mu}mol/g$ DW) under light condition and 'Mokpo No. 68' ($86.67{\mu}mol/g$ DW) in dark condition, whereas the lowest was in 'Tamra' (30.13 and $14.50{\mu}mol/g$ DW) in both conditions. The sum of aliphatic GSLs attributed > 80% in all varieties, except 'Tamra' (67.7% and 64.9% in dark and light conditions, respectively) in the total GSL accumulation. Indolyl GSLs were ranged $2.41-15.73{\mu}mol/g$ DW, accounted 2.78-33.6% of the total GSLs in rapeseed varieties. CONCLUSION(S): These results provide valuable information regarding potential beneficial GSL contents individually. This study attempts to contribute to knowledge of the nutritional properties of the different varieties of rapeseed plants. These results may be useful for the evaluation of dietary information.