Immuno-modulatory activities of peptides from Asterias amurensis were investigated using a nano-encapsulation process. The molecular weights of the peptides in the range of 5-7 kDa were separated using Sephadex G-75 gel filtration. Eighty-five percent of the nano-particles were in the 300 nm range using dynamic light scattering. The cytotoxicity of the A. amurensis nano-particles against CCD-986sk human dermal fibroblast cells was 11.64% after adding 1.0 mg/mL of the samples, which was lower than that from the control (13.28% collagen). The secretion of $NO^-$ from macrophages was estimated as $40\;{\mu}M$ after adding 1.0 mg/mL of gelatin nano-particles, which was higher than the others. Prostaglandin $E_2$ production from UV-induced human skin cells decreased greatly to 860 pg/mL after adding 1.0 mg/mL of the samples. Confocal microscopy revealed that nano-particles effectively penetrated the cells within 1 hour. From these results, we consider that nano-encapsulation of the peptides from A. amurensis can improve their biological functions.
Ha, Jun Young;Kim, Mi Kyeong;Lee, Jun Young;Choi, Eun Bi;Hong, Chang Oh;Lee, Byong Won;Bae, Chang Hwan;Kim, Keun Ki
Journal of Life Science
/
v.25
no.1
/
pp.53-61
/
2015
In this study, we tried to separate the photosensitizer that induces apoptosis of leukemia cells (U937) from perilla leaves. Perilla leaves (Perilla frutescens Britt var. japonica Hara) are a popular vegetable in Korea, being rich in vitamins (A and E), GABA, and minerals. Dried perilla leaves were extracted with methanol to separate the photosensitizer by various chromatographic techniques. The structure of the isolated compound (PL9443) was identified by 1D-NMR, 2D-NMR, and FAB-mass spectroscopy. Absorbance of the UV-Vis spectrum was highest at 410 nm and was confirmed by the 330, 410, and 668 nm. PL9443 compound was determined to be pheophorbide, an ethyl ester having a molecular weight of 620. It was identified as a derivative compound of pheophorbide structure when magnesium comes away from a porphyrin ring. Observation of morphological changes in U937 cells following cell death induced by treated PL9443 compound revealed representative phenomena of apoptosis only in light irradiation conditions (apoptotic body, vesicle formation). Results from examining the cytotoxicity of PL9443 substance against U937 cells showed that inhibition rates of the cell growth were 99.9% with the concentration of 0.32 nM PL9443. Also, the caspase-3/7 activity was 99% against U937 cells with the concentration of 0.08 nM of PL9443 substance. The result of the electrophoresis was that a DNA ladder was formed by the PL9443. The PL9443 compound is a promising lead compound as a photosensitizer for photodynamic therapy of cancer.
Lim, Jinkyu;Kim, Min-Yeol;Kim, Sung-Hee;Ma, Jin-Sung;Oh, Jisun;Kim, Jong Sang
Journal of Applied Biological Chemistry
/
v.60
no.4
/
pp.383-390
/
2017
Regarding the facts that fat, which is easily oxidized, is one of the major responsible factors affecting the quality of aroma, and polyphenol compounds including chlorogenic acid (CGA) contribute the anti-oxidative activities to coffee, we investigated fat oxidation, conversion of CGA, and changes of anti-oxidative activities according to the degree of roasting and storage of 60 days. We found that the amount of extractable fat by diethyl ether is increased as the coffee beans are roasted longer. Furthermore, the acidity values of the fat are increased from $8.91{\pm}0.16$ to $17.81{\pm}0.11$, and $10.37{\pm}0.27$ to $17.93{\pm}0.09$ in the medium and dark roasted coffee beans, respectively, while it is increased from $4.47{\pm}0.11$ to $11.89{\pm}0.18$ in the green coffee bean after 60 days. The CGA contents in the coffee beans were decreased from $310{\pm}8.2$ to $282{\pm}11.2$, then to $58{\pm}0.0mg$ in 10 gr of the green, medium and dark beans, respectively, and were not changed significantly during the storage period. However, the anti-oxidative activities measured by 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid radical scavenging assays were not significantly different among the green, medium, and dark coffee beans during the storage period. Furthermore, antioxidant reactive element-luciferase assay showed that biological anti-oxidative activities were increased as coffee beans were more roasted and stored longer. As the total polyphenolic contents in the beans were significantly decreased by roasting, the results suggests that other molecules, such as, Maillard reaction products might play substantial role in anti-oxidative activity and influence cup quality of coffee.
To study lipid components of Panax ginseng produced in Korea, the lipids of fresh ginsengs were extracted with the mixture of chloroform-methanol (2:1, v/v) and those of dried ginsengs were extracted with diethyl ether respectively. The lipid components extracted were separated and quantitated by column, thin layer and gas-liquid chromatographies. The results were summarized as follows : 1. Fresh ginseng contained 0.62% total lipid of which 45.28% were neutral lipids, 18.12% glycolipids, and 36.60% phospholipids. But dried ginseng contained 0.89% total lipids of which 86.48% were neutral lipids, 9.20% glycolipids, and 4.32% phospholipids. 2. Triglycerides (37.6 to 42.5% of the total neutral lipids) and sterol esters (16.5 to 19.6%) in all the fresh and dried ginseng were the major components among the neutral lipids. Monoglycerides, diglycerides, free fatty acids and free sterols were minor components. 3. Digalactosyl diglycerides (23.5% of the total glycolipids) in the fresh ginseng and steryl liglycosides (28.9%) in the dried ginseng were predominant components among the glycopids, respectively, Esterified steryl glycosides and monogalactosyl diglycerides were also identified, and four unknown spots in the fresh ginseng and two unknown spots in the dried ginseng were present. 4. Phosphatidyl cholines (31.3 to 31.9% of the total phospholipids) and phosphatidyl glycerols (34.8 to 36.7%) in all the fresh and dried ginseng were the major components among the phospholipids. Phosphatidyl inositols and phosphatidyl ethanolamines were also identified. 5. The major fatty acids in the fresh and dried ginseng were linoleic $(62.29{\sim}64.32%)$, palmitic $(13.16{\sim}15.63%)$, oleic $(5.73{sim}7.23%)$ and linolenic $(5.73{sim}7.23%)$. The fatty acid compositions in neutral lipid fraction was similar to the pattern in those of the total lipids. But glycolipid and phospholipid fractions contained a lower percent of linoleic acid and a higher percent of palmitic acid than the neutral lipid fraction.
Arsenic and its compounds vary in their toxicity according to the chemical forms. Inorganic arsenic is more toxic and known as carcinogen. The provisional tolerable weekly intake (PTWI) of $15{\mu}g/kg$ b.w./week established by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) has been withdrawn, while the EFSA panel suggested $BMDL_{0.1}$$0.3{\sim}8{\mu}g/kg\;b.w./day$ for cancers of the lung, skin and bladder, as well as skin lesions. Rice, seaweed and beverages are known as food being rich in inorganic arsenic. As(III) is the major form of inorganic arsenic in rice and anaerobic paddy soils, while most of inorganic arsenic in seaweed is present as As(V). The inorganic arsenic in food was extracted with solvent such as distilled water, methanol, nitric acid and so on in heat-assisted condition or at room temperature. Arsenic speciation analysis was based on ion-exchange chromatography and high-performance liquid chromatography equipped with atomic absorption spectrometry and inductively coupled plasma mass spectrometry. However, there has been no harmonized and standardized method for inorganic arsenic analysis internationally. The inorganic arsenic exposure from food has been estimated to range of $0.13{\sim}0.7{\mu}g/kg$ bw/day for European, American and Australian, and $0.22{\sim}5{\mu}g/kg$ bw/day for Asian. The maximum level (ML) for inorganic arsenic in food has established by EU, China, Australia and New Zealand, but are under review in Korea. Until now, several studies have conducted for reduction of inorganic arsenic in food. Inorganic arsenic levels in rice and seaweed were reduced by more polishing and washing, boiling and washing, respectively. Further research for international harmonization of analytical method, monitoring and risk assessment will be needed to strengthen safety management of inorganic arsenic of foods in Korea.
This study was conducted to analyze the microbial community and propionic acid production ability of natural microflora in the rice cakes. Genetic analysis of natural microflora in Jorangyi rice cake was performed to select propionic acid - producing bacteria. Selected propionic acid-producing bacteria were cultivated in TSB (tryptic soy broth) supplemented with glucose, and growth characteristics were analyzed by temperature and production of propionic acid was analyzed by gas chromatography (GC-FID). Linearity, detection limit, quantitative limit, and recovery rate were measured to verify propionic acid assay. A total of 98 microbial strains were detected from microflora of Joraengyi rice cake that grew after expiration of shelf life. Lactobacillus casei group accounted for 50.48% and Lactobacillus buchneri was 29.60%. Propionic acid - producing bacteria were Propionibacterium thoenii, P. cyclohexanicum, Propionibacterium_uc, P. jensenii, and P. freudenreichii. Natural bacteria and Lactobacillus spp. did not produce propionic acid during 14 days but P. cyclohexanicum, P. freudenreichii subsp. Shermanii, P. thoenii and P. jesenii produced $263.47{\mu}g/mL$, $338.90{\mu}g/mL$, $325.43{\mu}g/mL$ and $222.17{\mu}g/mL$ during 4 days and 2,462.02 and 2,904.78, 2,220.64, $3,519.17{\mu}g/mL$ during 14 days. As a result of this study, it was affirmed that the natural microflora of Joraengyi rice cake during storage can produce propionic acid from natural sources even if a high concentration of propionic acid is not intentionally added. Because of characteristics of rice cake composed of starch and glucose. This study will be used as a recognition criterion to detect natural preservatives such as propionic acid in starchy foods such as rice cakes and as reference standard safety management data.
Journal of the Korean Society of Marine Environment & Safety
/
v.13
no.1
s.28
/
pp.9-20
/
2007
PAHs(Polycyclic Aromatic Hydrocarbons) are widespread contaminants in the marine environment. They are of mainly anthropogenic origin from urban runoff, oil spill and combustion of fossil fuels. Some PAHs are potentially carcinogenic and mutagenic to aquatic organism The contamination of PAHs in the coastal environments has not been well known yet in Korea. This study was carried out to survey the contamination of PAHs in sediments around Gwangyang bay. The Yeosu petrochemical industrial complex, POSCO(Pohang steel company) and Gwangyang container harbor are located around the bay. PAHs in sediment samples were extracted in soxhlet extractor and were identified and quantified by GC-MS(Gas Chromatography-Mass Spectrometry) TOC(Total Organic carbon) and textural parameters in sediment samples were also analyzed 13 species of PAHs were detected at all of the surface sediments. Total PAHs concentrations in the surface sediments ranged from 171.40 to $1013.54{\mu}g/kg$ dry wt.. In most of the surface sediments, Naphthalene was the highest in the range of 14.08 to $691.39{\mu}g/kg$ dry wt. and Anthracene was the lowest in the range of 0.49 to $22.66{\mu}g/kg$ dry wt.. The correlation coefficients between individual PAHs and Total PAHs in the surface sediments were relatively higher in the low molecular compounds such as Naphthalene and Phenanthrene. In the relationship of the P/A(Phenanthrene/Anthracene) ratio and F/P(Fluoranthene/Pyrene) ratio, P/A ratio was generally above 10 and F/P ratio was shown to be above 1 in all sediment samples. These data indicate that PAHs in sediments around Gwangyang bay seem to be of both pyrolytic and petrogenic origin. Total PAHs in the surface sediments were correlated with TOC and textural parameters. The values of PAHs in the surface and core sediments were lower than the biological effect guidelines.
Lee, Jun Young;Kim, Mi Kyeong;Ha, Jun Young;Kim, Yong Gyun;Hong, Chang Oh;Kim, So Young;Kim, Chung-Hwan;Kim, Keun Ki
Journal of Life Science
/
v.24
no.3
/
pp.242-251
/
2014
The objective of this study was to isolate a photosensitizer from Pueraria thunbergiana leaves that induces apoptosis in SK-HEP-1 cells. Column chromatography and thin layer chromatography were used to isolate active compounds from extracts of P. thunbergiana leaves. The structures of the isolated compounds were determined by 1D-NMR, 2D-NMR, and FAB-mass spectroscopy. A substance, named M4-3, was purified from the leaves of P. thunbergiana using various chromatography methods, and the absorbance of the substance was measured. The absorbance was highest at 410 nm, suggesting that the M4-3 substance was a different compound from chlorophyll a and b, which absorb at 410, 502, 533, and 607 nm. Further analyses revealed that the M4-3 compound was a $13^2$-hydoxy pheophorbide, a methyl ester with a molecular weight of 662. M4-3 was identified as a derivative compound of pheophorbide, with a structure that magnesium comes away from the porphyrin ring. The results of the analysis of the cytotoxicity of the M4-3 substance against the SK-HEP-1 cells revealed that it inhibited rates of cell growth by 40% and 80% at a concentration of 0.04 ${\mu}M$ and 0.08 ${\mu}M$, respectively. The M4-3 compound was found to be a photosensitizer for cytotoxicity because it was appeared only in light condition as examining activity in different irradiation conditions (light condition and nonlight condition) under the same concentration. Analysis of morphological changes in the cells following cell death induced by exposure to the M4-3 substance reveled representative phenomena of apoptosis (nuclear condensation, vesicle formation, and fragmentation of DNA). The induction of apoptosis was attributed to the compound's photodynamic activity.
Park, Young-Il;Kim, Hee-Guen;Kim, Yoo-Young;Kim, In-Soo
Applied Biological Chemistry
/
v.39
no.6
/
pp.494-500
/
1996
Uptake of hen metal ions by water dropwort (Oenanthe stolonifera DC.) and its cadmium-binding protein were studied to probe for good method to remove heavy metal contaminants from environments. The plant was cultured in the culture medium (pH 7.0) containing the various concentrations of $Cd^{2+}$, $Cr^{3+}$ or $Pb^{2+}$, for 3 and 7 days. The residual heavy metals deposited in roots linearly increased as the metal ions concentration increased up to 17 ppm for $Cd^{2+}$, 20 ppm for $Cr^{3+}$ and 50 ppm for $Pb^{2+}$. Above these concentrations, the plant growth was inhibited and the uptake rates of the metal ions decreased. The heavy metals absorbed by the plant were mostly deposited in roots. In particular, the residual concentration of lead in roots was about four times higher than those of cadmium and chromium. When cultured in the medium containing 20 ppm of each metal ion, 80% of cadmium, 90% of cromium and 96% of lead were deposited in roots out of the total residual metal ions in the plant. These values correspond to 6.1 mg of cadmium, 5.2 mg of chromium and 23.6 mg of lead per one gram of roots tissue on a dry weight basis. A cadmium-binding protein was partially purified by extraction, gel filtration and DEAE-Cellulose chromatography from water dropworts that was grown in the medium containing 20 ppm $Cd^{2+}$. The purified protein was a single band on SDS- and non-denaturing- polyacrylamide gel electrophoresis. Its molecular mass was estimated to be ca. 5,000 dalton by gel filteration. Analysis of amino acid composition of the protein indicated that it had a typical amino acid composition of heavy metal-binding protein in that it contained 27% of acidic amino acids and 9.9% of cysteine. However, it is likely that the protein is a new plant metal-binding protein, since its amino acid composition is somewhat different from those of phytochelatins that have been known so far.
YANG Huyn-Pil;LEE An-Jong;KIM Yong-Tae;KIM Se-Kwon
Korean Journal of Fisheries and Aquatic Sciences
/
v.27
no.5
/
pp.482-494
/
1994
Most of carotenoprotein complexes have been extracted by using buffered solutions. However, in this study carotenoprotein from the muscle of Blue mussel(Mytilus edulis) was extracted by a detergent such as Triton X-100. It was purified and characterized by $20\%$ (w/v) $(NH_4)_2SO_4$, DEAE-cellulose ion exchange and Sephacryl S-300 gel filtration. The carotenoprotein(${\lambda}_{max}=462nm$) had an approximate M. W. of 372KDa(gel filtration). SDS-PAGE analysis of the carotenoprotein indicated the presence of four polypeptides of 60KDa($23.70\%$), 46.9KDa($9.14\%$), 26KDa($49.14\%$) and 13KDa($18.02\%$). Carotenoprotein denaturated by treatment with SDS to a final concentration of $0.2\%$ (w/v) caused a hypsochromic shift of ${\lambda}_{max}$ from 462nm to 456nm. The carotenoprotein contained lipids as structure units. The amino acid composition of the carotenoprotein contained large essential amino acid amounts of $62.8\%$, and the content of threonine($35.9\%$) was higher than other amino acids, but histidine, methionine and proline were not present. In the carotenoprotein, the major fatty acids were $C_{16:4},\;C_{16:0},\;C_{20:5}\;and\;C_{22:6}$. The percentages of polyunsaturated fatty acids($62.4\%$) were higher compared to other fatty acids(saturated fatty acids $19.6\%$, monounsaturated fatty acids $18.0\%$). Carotenoid was extracted from the carotenoprotein by acetone and it was separated into five different components by preparative TLC(benzene:petroleum ether:acetone=69:17:14). The major components of carotenoid were mytiloxanthin($74.79\%$) and 3,4,3'- trihydroxy-7',8'-didehydro-${\beta}$-carotene($18.26\%$), and they were at least presented as prosthetic groups of carotenoprotein.
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