• Journal of Microbiology and Biotechnology
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• 제14권5호
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• pp.985-990
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• 2004

Levans produced from Microbacterium laevaniformans were isolated, characterized, and fractionated by molecular weight. TLC, HPLC, and GC-MS analyses of the exopolysaccharide showed that it was a fructan-type polymer and was composed of (2,6)- and (2,1)-glycosidic linkages. $^{13}C$-NMR analysis proved that the polysaccharide was mainly a $\beta$-(2,6)-linked levan-type polysaccharide. To investigate the cytotoxicity of the acetone-precipitated levan fractions such as M1, M2, and M3, HepG2, P388D1, U937, SNU-1, and SNUC2A cell lines were screened. Among the cell lines tested, the cytotoxicity of M1- M3 fractions were detected from only SNU-1 and HepG2 cells at the dosage level of $100-800\mu\textrm{g}ml$. The M2 fraction M_r$, 80,000) at 400 $mu{g/ml}$ had the greatest cell growth inhibition (84.6%) on SNU-1, while the M1 $(M_r$, 50,000) at $800\mu\textrm{g}ml$ showed the greatest (46.32%) on HepG2. To obtain more uniform M_r$ fractions of levan, the levan was further fractionated from S1 $(M_r$ 1,000,000) to S5 $(M_r$ 10,000) using gel permeation chromatography. Again, the S1-S5 fractions had strong cytotoxicity on SNU-1 and HepG2 cell lines. The greatest inhibition effects of S4 $(M_r$ 80,000) on SNU-1 and S5 $(M_r$ 10,000) on HepG2 were shown to be 49.5% and 73.0%, respectively. The cytotoxicity of the levan fractions was more effective on SNU-1 than on HepG2. Although the relationship between the Mw and the cytotoxicity was not clear, smaller $M_r$, fractions of levan showed greater growth inhibition effect on the cancer cell lines in general. Therefore, it was indicated that a specific Mw class of levan is responsible for the effective cytotoxicity.

• Applied Biological Chemistry
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• 제37권1호
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• pp.43-48
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• 1994

A basic inducible protein, ICG, containing chitinase and ${\beta}-1,3-glucanase$ activity concomittantly was purified from cell suspension culture media of rice after the treatment of chitooligosaccharides. The isolated ICG enzyme gave a single band on native and SDS polyacrylamide gel electrophoresis and its molecular weight was estimated to be 52.53 kd. The optimal temperature and optimal pH of both enzyme activities in ICG were $60^{\circ}C$, pH 6.0 for chitinase activity and $37^{\circ}C$, pH 4.0 for ${\beta}-1,3-glucanase$ activity. $K_M$ and $V_{max}$ values for chitinase were 0.474 mM. 2.997 nM/min., and those for ${\beta}-1,3-glucanase$ were 1.004 mM 0.739 nM/min. respectively. TLC analysis of the chitooligosaccharide hydrolysates with ICG enzyme indicated that ICG acts as endochitinase.

• Asian-Australasian Journal of Animal Sciences
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• 제20권7호
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• pp.1067-1075
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• 2007

Twenty eight 3-4 month old castrated Black Bengal kids (Capra hircus) were used to determine the effects of source and level of dietary copper (Cu) concentration on their performance and nutrient utilization. Cu was supplemented (0, 10, 20 and 30 mg/kg diet DM) as copper sulfate ($CuSO_4$, $5H_2O$) or copper proteinate (Cu-P). Kids were fed a basal diet containing maize (19.5%), soybean (17.0%), deoiled rice bran (56.5%), molasses (4.0%), di-calcium phosphate and salt (1.0% each) and mineral and vitamin mixture (0.5% each) supplements at 3.5% of body weight to meet NRC (1981) requirements for protein, energy, macro minerals and micro minerals, excluding Cu. The basal diet contained 5.7 mg Cu/kg, 122.5 mg Fe/kg, 110 mg Zn/kg, 0.26 mg Mo/kg and 0.32% S. $CuSO_4$ or Cu-P was added to the basal diet at the rate of 10, 20 and 30 mg/kg. Kids were housed in a well ventilated shed with facilities for individual feeding in aluminum plated metabolic cages. Blood samples were collected from the jugular vein on d 0, 30, 60 and 90 to determine hemoglobin (Hb), packed cell volume (PCV), total erythrocyte count (TEC), total leukocyte count (TLC) and serum enzymes (alkaline phosphatase, alanine transferase and aspertate transferase). A metabolism trial of 6 days duration was conducted after 90 days of experimental feeding. Statistical analysis revealed that source and level of Cu supplementation improved live weight gain (p<0.04) and average daily gain (p<0.01). No significant contribution of source and level of Cu to alter serum serum enzymes was evident. Goats fed Cu-P tended to have higher Hb, PCV and TEC than with $CuSO_4$ supplementation. Cu-P increased digestibility of ether extract (EE, p<0.02) and crude fiber (p<0.05) and showed an increasing trend (p<0.09) for digested crude protein (CP) and crude fiber (CF). Supplemental dose of Cu linearly improved (p<0.02) digestibilities of dry matter (DM), organic matter (OM), EE and nitrogen free extract (NFE). Though the absorption of nitrogen (N) was not affected (p>0.10) by both source and dose of Cu, N retention was affected (p<0.04) and there was a significant $Source{\times}Dose$ interaction (p<0.05). Final body weight (BW) was not influenced (p>0.10) by the source of Cu but increasing dose of Cu increased (p<0.04) the BW of kids. TDN intake (g/kg $W^{0.75}$) was higher (p<0.05) with the increased dose of Cu and there was a significant $Source{\times}Dose$ interaction. It was concluded that supplementation of Cu from different sources and varying dose level in a concentrate based diet may improve performance, nutrient utilization and plane of nutrition in castrated Black Bengal kids. The effects on performance and nutrient utilization are more pronounced with Cu-P than $CuSO_4$ supplementation. Higher dose of Cu showed better result than lower dose.

• Journal of Microbiology and Biotechnology
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• 제10권1호
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• pp.27-34
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• 2000

The chloroform and methanol (2;1, v/v) extract from an edible plant, Actinidia arguta Planchon, appeared to possess antitumor activity against human leukemias Jurkat T and U937 cells through inducing apoptosis. The substance in the solvent extract was purified by silica gel column chromatography, preparative TLC, and Sephadex LH-20 column chromatography. Characteristics of the substance analyzed by UV scanning analysis, $^1H$ and $^{13}C$ NMR spectra suggested that the substance belongs to the chlorophyll derivatives-like group. The $IC_{50}$ value of the chlorophyll derivative (Cp-D) determined by MTT assay was $15\mu\textrm{g}/ml$ for Jurkat, $10\mu\textrm{g}/ml$ for U937, and $11.4\mu\textrm{g}/ml$ for HL-60m and was more toxic to these leukemias than to solid tumors or normal fibroblast. In order to elucidate cellular mechanisms underlying the cytotoxicity, the effect of the Cp-D on Jurkat T cells was investigated. When cells were treated with the Cp-D at a concentration of $15\mu\textrm{g}/ml$, [3H]thymidine incorporation declined rapidly and wa undetectable in 1h. However, no significant changes were made in the cell cycle distribution of the cells by 24h. The sub-Gl peak representing apoptotic cells began to be detectable in 36h, at which time apoptotic DNA fragmentation was also detected on agarose gel electrophoresis, demonstrating that the cytotoxic effect of the Cp-D is attributable to the induced apoptosis. Under the same conditions, although the protein level of cyclin-dependent kinases such as cdc4, csk6, cdk2, and cdc2 was not significantly changed until 24h, the kinase activity of all c안 rapidly declined and reached a minimum level within 1-6h and then recovered to the initial level by 12h and sustained until 24h. These results suggest that inactivation of cdks at an inappropriate time during the cell cycle progression in jurkat T cells following a treatment with the Cp-D leads to induction of apoptotic cell death.

• Journal of the Korean Wood Science and Technology
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• 제38권4호
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• pp.359-374
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• 2010

This study was carried out to investigate the chemotaxonomical correlation and chemical constituents of domestic Quercus spp. barks. The barks of Q. mongolica, Q. aliena, Q. serrata, Q. acutissima, Q. dentata, and Q. variabilis were collected in the experimental forest of Kangwon National University. The combined extracts were successively fractionated with n-hexane, methylene chloride and ethyl acetate using a separation funnel. A portion of the ethyl acetate and H2O soluble materials of each species were chromatographed on a Sephadex LH-20 column using various aqueous MeOH and EtOH-hexane as washing solvents. Spectrometric analysis such as NMR and MS, including TLC, were performed to characterize the structures of the isolated compounds. Ellagic acid (0.03 g), (+)-catechin (4.59 g), taxifolin (3.35 g), and glucodistylin (20.52 g) were isolated from Q. mongolica bark. Gallic acid (0.18 g), (+)-catechin (8.52 g), (+)-gallocatechin (0.09 g), taxifolin (0.54 g), and glucodistylin (3.28 g) were characterized from Q. acutissima bark. Gallic acid (0.38 g), ellagic acid (0.11 g), (+)-catechin (2.01 g), (+)-gallocatechin (0.12 g), and glucodistylin (0.39 g) were identified from Q. dentata bark. Ellagic acid (1.51 g), (+)-catechin (21.91 g), and glucodistylin (3.91 g) were purified from Q. aliena bark. Ellagic acid (0.84 g), (+)-catechin (0.82 g), taxifolin (4.02 g), and glucodistylin (21.50) were isolated from Q. serrata bark. Gallic acid (0.24 g), caffeic acid (0.05 g), (+)-catechin (0.32 g), and glucodistylin (0.65 g) were purified from Q. variabilis bark. (+)-Catechin and glucodistylin were isolated from all the barks. Glucodistylin can be a taxonomic index on Quercus spp.

• Analytical Science and Technology
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• 제13권2호
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• pp.136-150
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• 2000

For the identification of unknown drugs in biological samples, we attempted rapid high performance thin layer chromatographic method which is sensitive and selective chromatographic analysis of high performance thin layer chromatography (HPTLC) with automated TLC sampler and ultra-violet (UV) scanner. We constructed HPTLC database (DB) on two hundred five drugs by using the data of Rf values and UV spectra (scan 200-360 nm) as well as gas chromatography/mass spectrometry (GC/MS) DB on ninety six drugs by using the data of relative retention time (RRT) on lidocain and mass spectra. After extracting drugs in biological sample by solid phase extraction (Clean Screen ZSDAU020), we applied them to HPTLC and GC/MS DB. Drugs, especially extracted from biological samples, showed good matching ratio to HPTLC DB and these drugs were confirmed by GC/MS. In conclusion, this DB system is thought to be very useful method for the screening of unknown drugs in biological samples.

• Korean journal of food and cookery science
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• 제15권6호
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• pp.645-651
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• 1999

The lipid and fatty acid composition of various poultry eggs were examined after extracting yolk oils from the eggs of caged hen, open barn-reared hen, mallard, and muscovy duck. Total lipid content in 100 g of each edible portion was the highest in muscovy duck egg followed by open barn-reared hen, mallard, caged hen, and natural hen eggs. The triglyceride contents in the egg were in the order of caged hen, open barn-reared hen, natural hen, mallard, and muscovy duck. The cholesterol contents in total lipid were the highest in caged hen and muscovy duck eggs(4.5%), and the lowest in mallard(3.3%). Caged hen eggs had higher neutral lipid ratio among total lipid, but had lower ratio of glycolipid and phospholipid compared with those of the open barn-reared hens. For fatty acid composition, linoleic acid was the highest in the muscovy and linolenic acid was the highest in open barn-reared hen eggs. The contents of arachidonic acid and Eicosapentaenoic acid(EPA) were the highest in muscovy eggs. On the other hand, the content of Docosahexaenoic acid(DHA) was the highest in muscovy duck eggs.

• Korean Journal of Community Nutrition
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• 제13권1호
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• pp.80-90
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• 2008

Atopic dennatitis (AD) is one of the major public health problem. It has been reported that the prevalence of AD in children and adults are 10-20% and 1-3%, respectively. Westernization of food habits, urbanization, and environmental pollution are contributing factors toward the recent rise in prevalence. Excessive dietary restriction leads to chronic malnutrition in atopic dermatitis patients. The purpose of this study was to investigate the effects of medical nutrition therapy (MNT) on quality of diet and blood immune parameters in atopic dermatitis patients. The 19 atopic dermatitis patients (7 men and 12 women) admitted to K University Medical Center were studied. During the 12 weeks of intervention, the subjects were given MNT by a dietitian for 30-45 minutes every other week. MNT was comprised with general dietary therapy, intake of balanced meals, emphasis on n-3 fatty acid contents in foods, and food allergies. Anthropometric and dietary assessment and blood analysis were taken at baseline and after 12 weeks of MNT. After 12 weeks of MNT, the subjects' dietary qualities, including dietary diversity score (DDS), meal balance score (MBS) and dietary variety score (DVS) were significantly increased (p < 0.05). According to significantly increased intake of EPA and DHA, dietary n-6/n-3 fatty acid ratio decreased to the recommended level for the atopic dermatitis patients (p < 0.05). These changes of dietary fatty acid consumption were reflected erythrocyte fatty acid composition. After 12 weeks of MNT, serum levels of IgE and IL-4 levels were significantly decreased, however, the levers of INF-$\{gamma}$, WBC, lymphocyte and TLC were not changed. As a conclusion, the individualized MNT improved the quality of diet in atopic dermatitis patients thereby influenced RBC fatty acid composition and IgE and IL-4 levels.

• Journal of the Korea Academia-Industrial cooperation Society
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• 제16권11호
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• pp.7794-7800
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• 2015

Ginsenosides in ginseng berry has been known as functional materials showing physiological effect to the human. Specially, ginseng berry contains plenty of ginsenoside Re, but the study of extraction processes were not enough performed. Accordingly, the purpose of this study was to establish the optimized extraction process for obtaining ginsenoside Re from ginseng berry. The extraction process of ginsenosides was performed in 250 mL extraction flask containing 150 solvent and 10 g of dried ginseng berry. The extracted ginsenoside Re, Rg1 and Rd and total crude ginsenosides from ginseng berry were evaluated by TLC according to the treated conditions (the ratio of alcohol to water, extraction temperature, extraction period, and extraction times). Optimized conditions for extraction was 70% to 30% of the ratio of alcohol to water, $80^{\circ}C$ of extraction temperature, 4 h of extraction period, and 2 times of extraction frequency. The amount of total crude ginsenosides of the extract obtained from the optimized process was 88.6 mg/g based on dried ginseng berry. The composition of ginsenosides from the extracted was 5.5% of Rb1, 5.2% of Rc, 14.3% of Rd, 51.5% of Re, 8.1% of Rf, and 15.7% of Rg1. A protopanaxtriol ginsenosides of whole ginsenosides extracted was about 80%.

• Journal of Life Science
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• 제28권9호
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• pp.1056-1061
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• 2018

This article reports an agar-degrading marine bacterium and characterizes its agarase. The agar-degrading marine bacterium, KC-1, was isolated from seawater on the shores of Sacheon, in Gyeongnam province, Korea, using Marine Broth 2216 agar medium. To identify the agar-degrading bacterium as Agarivorans sp. KC-1, phylogenetic analysis based on the 16S rRNA gene sequence was used. An extracellular agarase was prepared from a culture medium of Agarivorans sp. KC-1, and used for the characterization of enzyme. The relative activities at 20, 30, 40, 50, 60, and $70^{\circ}C$ were 65, 91, 96, 100, 77, and 35%, respectively. The relative activities at pH 5, 6, 7, and 8 were 93, 100, 87, and 82%, respectively. The extracellular agarase showed maximum activity (254 units/l) at pH 6.0 and $50^{\circ}C$ in 20 mM of Tris-HCl buffer. The agarase activity was maintained at 90% or more until 2 hr exposure at $20^{\circ}C$, $30^{\circ}C$ and $40^{\circ}C$, but it was found that the activity decreased sharply from $60^{\circ}C$. A zymogram analysis showed that Agarivorans sp. KC-1 produced 3 agar-degrading enzymes that had molecular weights of 130, 80, and 69 kDa. A thin layer chromatography analysis suggested that Agarivorans sp. KC-1 produced extracellular ${\beta}$-agarases as it hydrolyzed agarose to produce neoagarooligosaccharides, including neoagarohexaose (21.6%), neoagarotetraose (32.2%), and neoagarobiose (46.2%). These results suggest that Agarivorans sp. KC-1 and its thermotolerant ${\beta}$-agarase would be useful for the production of neoagarooligosaccharides that inhibit bacterial growth and delay starch degradation.