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The Screening of Nitrite Scavenging Effect of Marine, Algae and Active Principles of Ecklonia Stolonifera

  • CHOI Jae Sue;LEE Ji Hyeon;JUNG Jee Hyung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.6
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    • pp.909-915
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    • 1997
  • The nitrite scavenging effect of methanol extracts of marine algae were evaluated to discover new natural nitrite scavengers. Among the tested seaweeds, Ecklonia stolonifera, an edible brown algae, showed the strongest scavenging effect. The MeOH extract was then sequentially partitioned into $CH_2Cl_2,\;CH_2Cl_2$ insoluble interface, EtOAc, n-BuOH, and $H_2O$ layers. The EtOAc and n-BuOH fraction demonstrated high levels of nitrite-scavenging activity while the $CH_2Cl_2,\;CH_2Cl_2$ insoluble interface, and $H_2O$ fractions were inactive. A column chromatography of the EtOAc fraction through silica gel and Sephadex LH-20 yielded phloroglucinol and a new compound tentatively named phlorotannin A. The nitrite scavenging activity of phloroglucinol $(IC_{50}=3.9{\mu}g/ml)$ was more potent than that of L-ascorbic acid $(IC_{50}=65.0{\mu}g/ml)$. However, phlorotannin A $(IC_{50}=193.2{\mu}g/ml)$ showed only low levels of activity. From the above results, it is possible to suggest that both the MeOH extract and their fractions and isolated phloroglucinol and phlorotannin A obtained from E. stolonifera may be applicable as scavengers of nitrite, which is a precursor for the formation of carcinogenic N-nitroso compounds.

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Bioproduction and Anticancer Activity of Biosurfactant Produced by the Dematiaceous Fungus Exophiala dermatitidis SK80

  • Chiewpattanakul, Paramaporn;Phonnok, Sirinet;Durand, Alain;Marie, Emmanuelle;Thanomsub, Benjamas Wongsatayanon
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1664-1671
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    • 2010
  • A new biosurfactant producer was isolated from palm-oil-contaminated soil and later identified through morphology and DNA sequencing as the yeast-like fungus Exophiala dermatitidis. Biosurfactant production was catalyzed by vegetable oil, supplemented with a basal medium. The culture conditions that provided the biosurfactant with the highest surface activity were found to be 5% palm oil with 0.08% $NH_4NO_3$, at a pH of 5.3, with shaking at 200 rpm, and a temperature of $30^{\circ}C$ for a 14-day period of incubation. The biosurfactant was purified, in accordance with surfactant properties, by solvent fractionation using silica gel column chromatography. The chemical structure of the strongest surface-active compound was elucidated through the use of NMR and mass spectroscopy, and noted to be monoolein, which then went on to demonstrate antiproliferative activity against cervical cancer (HeLa) and leukemia (U937) cell lines in a dose-dependent manner. Interestingly, no cytotoxicity was observed with normal cells even when high concentrations were used. Cell and DNA morphological changes, in both cancer cell lines, were observed to be cell shrinkage, membrane blebbling, and DNA fragmentation.

Glioblastoma-Specific Anticancer Activity of Pheophorbide a from the Edible Red Seaweed Grateloupia elliptica

  • Cho, MyoungLae;Park, Gab-Man;Kim, Su-Nam;Amna, Touseef;Lee, Seokjoon;Shin, Woon-Seob
    • Journal of Microbiology and Biotechnology
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    • v.24 no.3
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    • pp.346-353
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    • 2014
  • The chlorophyll-related compound pheophorbide a (Pa) was successively purified from an edible red seaweed, Grateloupia elliptica, using silica, octadecyl silica column chromatography and reversed phase-high-performance liquid chromatography, as well as the cell cycle inhibitory and apoptotic effects of Pa being investigated in U87MG glioblastoma cells. The Pa exhibited strong anticancer effects in the absence of direct photo-irradiation against various cancer cell lines, including U87MG, SK-OV-3, and HeLa cells. Among the cancer cells, the strongest anticancer activity of Pa exhibited on U87MG cells with $IC_{50}$ values of 2.8 ${\mu}g/ml$. In addition, Pa specifically had cytostatic activity on glioblastoma cells rather than human umbilical vein endothelial cells. Analysis of the cell cycle distribution showed that Pa induced G0/G1 arrest of U87 MG cells. In addition, arrested cells induced late apoptosis and DNA degradation under dark condition. These results suggest that Pa isolated from G. elliptica is a potential glioblastoma-specific anticancer agent without side effects on normal cells.

Screening for Antioxidant Activity of Plants and Marine Algae and Its Active Principles from Prunus davidiana

  • Choi, Jae-Sue;Lee, Ji-Hyeon;Park, Hae-Jin;Kim, Hyung-Geun;Young, Han-Suk;Mun, Sook-Im
    • Korean Journal of Pharmacognosy
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    • v.24 no.4
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    • pp.299-303
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    • 1993
  • The antioxidant activity of methanol extracts of plants and marine algae was tested by using 1,1-diphenyl-2-picrylhydrazyl(DPPH). Five plant extracts(Prunus davidiana, Eriobotrya japonica, Artemisia iwayomogi, Stirodella tolyrrhiza and Ulmus davidiana) and two algae (Ecklonia stolonifera and Symphycoladia latiuscula) were found to be the most effective in DPPH radical scavenging activity. The methanol extract obtained from the stems of Prunus davidiana was fractionated with several solvents. The ethylacetate soluble fraction exhibiting the strongest antioxidant activity was further purified by repeated silica gel and Sephadex LH-20 column chromatography. Antioxidant flavonoids and flavonoid glycosides were isolated and the most active ones was identified as (+)-catechin by MS, $^1H-NMR$ and $^{13}C-NMR$. Its antioxidant activity was higher than that oil vitamin C.

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Isolation and Characterization of Antioxidative Peptides from Enzymatic Hydrolysates of Yellowfin Sole Skin Gelatin (가자미피 젤라틴 가수분해물로부터 항산화성 펩티드의 분리${\cdot}$정제 및 특성)

  • KIM Se-Kwon;LEE Hyun-Chel;BYUN He-Guk;JEON Yon-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.2
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    • pp.246-255
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    • 1996
  • To develop a natural antioxidative peptide, the gelatin was extracted from fish (Yellowfin sole) skin by hot $water(50^{\circ}C)$ extraction method and hydrolyzed with Alcalase, pronase and collagenase through a continuous 3-step membrane reactor. Each step enzymatic hydrolysates were determined the antioxidative activity and their synergistic effects, compared with $\alpha-tocopherol$ and butylated hydroxytoluene (BHT). Also, we tried to investigate the antioxidative disposition of peptide which was successfully separated by gel filtration, ion-exchange chromatography, and HPIC in cultured rat hepatocytes intoxicated with tert-butyl hydroperoxide (TBHP). Second step enzymatic hydrolysate (SSEH) among all hydrolysates and $\alpha-tocoperol$ was showed the strongest antioxidative activity. The optimum concentration of antioxidative activity for SSEH was $1\%(w/w)$ in linoleic acid. The synergistic effects were increased in using the hydrolysate with tocopherol and BHT. In the presence of the peptide isolated from SSEH, supplemented hepatocytes exposed to TBHP showed that delayed cell killing and decreased significantly the lipid peroxidation, compared with hepatocytes not cultured with isolated peptide.

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Purification and Characterization of Antioxidative Peptides from Enzymatic Hydrolysate of Cod Teiset Protein (대구고니 단백질의 효소적 가수분해물로부터 항산화성 펩타이드의 분리${\cdot}$정제 및 특성)

  • KIM Se-Kwon;CHOI Yong-Ri;PARK Pyo-Jam;CHOI Jeoung-Ho;MOON Sung-Hoon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.3
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    • pp.198-204
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    • 2000
  • In order to utilize by-products which would normally be discarded in marine processing plants, cod teiset protein was hydrolyzed and antioxidative actiTity of the hydrolysate was investigated. AntioxidatiTe peptide was isolated using ultrafiltration membrane, ion-exchange chromatography on a SP-Sephadex C-25 column, gel filtration on a Sephadex G-15 column, high performance liquid chromatography on an ODS column, and capillary electrophoresis chromatography. Antioxidative activities of the cod teiset hydrolysate were compared with ${\alpha}-tocopherol$, one of the commercial antioxidant. The hydrolysate passed through a membrane with molecular weight cut-off (MWCO) 1 kDa was shown the strongest antioxidative activity, and the activity was higher $10{\%}$ as compared with ${\alpha}-tocopherol$. In addition, the peptide isolated by ion-exchange chromatography, gel filtration, and HPLC, respectively, was higher $53{\%}$ as compared with ${\alpha}-tocopherol$, and the amino acid sequence was Ser-Asn-Pro-Glu-Trp-Ser-Trp-Asn.

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Lipase Inhibitory Mode of Dieckol Isolated from Eisenia bicyclis Ethanol Extract (Eisenia bicyclis 에탄올 추출물로부터 분리한 Dieckol의 Lipase 저해 Mode)

  • Jung, Seul-A;Kim, Koth-Bong-Woo-Ri;Kim, Dong-Hyun;Cho, Ji-Young;Kim, Tae-Wan;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.112-118
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    • 2013
  • This study was performed to investigate the possible use of Eisenia bicyclis (EB) ethanol extract to inhibit activity against lipase. In tests, the lipase inhibitory activity of EB ethanol extract was noted as being 43, 27, and 24% at concentrations of 5, 2.5, and 1 mg/ml, respectively. Isolation was carried out by liquid and liquid extraction, silica-gel column chromatography, and HPLC. The results showed that the lipase inhibitory activity of the ethyl acetate (EA) fraction from EB ethanol extract exhibited the strongest lipase inhibitory activity with an $IC_{50}$ value of 1.31 mg/ml. The EA fraction was separated using silica-gel column chromatography and we obtained 22 sub-fractions. Amongst them, the EA1 fraction showed the highest lipase inhibitory activity with an $IC_{50}$ value of 0.54 mg/ml. Eight peaks were obtained from the EA1 fraction by HPLC. Fraction 5 also showed a strong lipase inhibitory activity with an $IC_{50}$ value of 0.37 mg/ml. The fraction 5 was identified as dieckol and the inhibition pattern analyzed from Lineweaver-Burk plots revealed a non-competitive inhibitor. These results suggest that EB has potential as a natural anti-obesity agent.

Characteristics of Nitrite Scavenger Derived from Seeds of Cassia tora L. (결명자 추출물의 아질산염 소거인자의 특성)

  • Park, Yeung-Beom;Lee, Tae-Gee;Kim, Oi-Kyung;Do, Jeong-Ryong;Yeo, Saeng-Gyu;Park, Yeung-Ho;Kim, Seon-Bong
    • Korean Journal of Food Science and Technology
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    • v.27 no.1
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    • pp.124-128
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    • 1995
  • The present study was conducted to elucidate nitrite scavenger derived from seeds of Cassia tora L. Methanolic extract of Cassia tora L was refractionated into ethyl ether, chloroform, ethyl acetate and water farction, and nitrite scavenging abilities of these fractions were investigated. Among these fractions, ethyl acetate fraction had the strongest nitrite scavenging ability(97%/2 mg). Therefore, to further investigate nitrite scavenger, ethyl acetate fraction was fractionated by silica gel column chromatography with a chloroform-methanol($10:0{\sim}0:10$) and then compound A and B were isolated. Compound A had a stronger nitrite-scavenging effect than compound B. Therefore the further separation of compound A was carried out by HPLC(32% acetonitrile. 1 ml/min) using ${\mu}Bondapak$C18 column$(3.9{\times}300 mm)$ and finally compound A-1 was obtained from compound A. Compound A-1 had by far nitrite-scavenging ability as compared with that of ascorbic acid. Compound A-1 was identified as $nor-rubrofusarin-6-{\beta}-mono-D-glucoside$ from the profiles of UV, IR and $^1H-NMR$.

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Isolation and Identification of Antioxidant Compounds of Various Solvents Extracted from Eriobotrya japonica Leaves (비파 잎의 추출용매별 항산화성 검증과 활성물질의 분리 및 동정)

  • Ham, Hyeon-Suk;Lee, Se-Yeul;Lee, Dong-Wan;Seong, Jong-Hwan;Kim, Han-Soo;Kim, Dong-Seob;Lee, Young-Guen
    • Journal of Life Science
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    • v.22 no.9
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    • pp.1166-1172
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    • 2012
  • To investigate potential medicinal or functional uses of Eriobotrya japonica, this study focused on the isolation and identification of antioxidant compounds from Eriobotrya japonica leaves. Various solvents were extracted from the leaves, and their scavenging effect on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals was measured, in addition to their superoxide dismutase-like activity, polyphenol compounds, and flavonoid content. Ethyl acetate extract exhibited the strongest scavenging effect in a 0.2 mM solution of DPPH ($63.24{\pm}2.20%$, $81.83{\pm}2.10%$, and $93.15{\pm}2.31%$ in 0.3, 0.7, and 1.0 mg/ml sample concentrations, respectively). The antioxidant effect of the ethyl acetate extract and methanol extract were generally stronger than that of n-hexane extract. The extracts were further purified by repeated silica gel column chromatography. The antioxidant compounds were identified as phytol, ${\beta}$-sitosterol, and (-)-loliolide using GC/MS.

Antioxidant Activities of Stachys sieboldii MIQ. Stalks (초석잠 [Stachys sieboldii MIQ.] 줄기의 항산화 활성)

  • 백홍석;나영수;류병호;송승구
    • KSBB Journal
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    • v.18 no.4
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    • pp.266-271
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    • 2003
  • This study was worked out to investigate the compounds of antioxidant constituents extracted from Stachys Sieboldii MiQ. and their effects on antioxidant activity by DPPH method, ferric thiocyanate method, and nitrite scavenging ability. Solvents such as methanol, hexane, chloroform, ethyl acetate, butanol, and water were used for this purpose. Total concentrations of polyphenols and flavonoids were measured in the methanol fraction. The ethyl acetate fraction showed the strongest activity by DPPH method, ferric thiocyanate method, and nitrite scavenging ability. The ethyl acetate extract was fractionated on a silica gel column using elution solvent (chloroform: methanol: water = 70 : 30 : 5 lower phase) at a flow rate of 1.0 mL/min. UV-VIS spectral data of each fraction showed adsorption maxima in the range of 284~330 nm. Among fractions, the fraction 1 that has λ$\_$max/ (nm) of 284 nm showed the strongest activity by DPPH method. The UV-VIS spectral data of phenolic compounds were known to lie in the range of 210~290 nm and 300~550 nm. Therefore, the results of our study suggested that Stachys sieboldii MIQ. contains phenolic compounds showing natural antioxidant activity.