• Journal of Applied Biological Chemistry
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• v.52 no.4
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• pp.180-186
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• 2009

Whole plants of Vitex rotundifolia were extracted for 2 days with methylene chloride ($CH_2Cl_2$) followed by extraction of the residue for an additional 2 days. The same procedure was also applied using methanol (MeOH). The two crude extracts were combined and partitioned between $CH_2Cl_2$ and $H_2O$. The organic layer was further partitioned between n-hexane and 85% aq. MeOH, and the aqueous layer was also further fractionated with n-BuOH and $H_2O$, successively. From the 85% aq. MeOH fraction, one compound was isolated through the repeated HPLC. According to the results of physicochemical data including NMR and MS, the chemical structure of the compound was determined as artemetin (1). The antiproliferative effects of the crude extracts, fractions, and compound against HT1080, AGS, MCF-7 and HT-29 human cancer cells were compared with the control by using MTT assay. In the comparative analysis, the 85% aq. MeOH fraction exhibited the strongest antiproliferative effects on human cancer cell lines in a dose-dependent manner (p<0.05). In addition, exposure of compound 1 isolated from 85% aq. MeOH fraction led to strong antiproliferative effect in HT1080 cancer cell lines. These results suggest that the extracts and compound isolated from V. rotundifolia may be used as potential chemopreventive and chemotherapeutic agents.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.4
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• pp.395-401
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• 2006

In this study, we investigated anticarcinogenic effects of extracts from Gloiopeltis tenax (GT). GT was extracted with methanol (GTM), which was then further fractionated into four fractions by using solvent fractionation method, affording methanol (GTMM), hexane (GTMH), butanol (GTMB) and aqueous (GTMA) soluble fractions. We determined the cytotoxic effects of these fractions on cancer cells by MTT assay. Among various fractions of GT, the GTMM showed the strongest cytotoxic effect at concentration of $150{\mu}g/mL$, displaying 95.97% on HepG2 cell lines and 93.64% on HT-29 cell lines, respectively. And, the anti-proliferative effect of GT was accompanied by a marked in increase of levels of Bad, Bax, Bok and Bak protein and activation of caspase-3, caspase-7 and PARP protein. Also, we observed quinone reductase (QR) induced effects in all fraction layers of GT on HepG2 cells. The QR induced effects of the GTMM and GTMB on HepG2 cells at concentration of $60{\mu}g/mL$ showing inductive indexes of 2.86 and 2.04 compared to the control value of 1.0.

• KSBB Journal
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• v.23 no.2
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• pp.177-182
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• 2008

In this study, crude extracts of the halophyte Limonium tetragonum and their solvent fractions were evaluated on anticancer activity in AGS and HT-29 human cancer cells using MTT assay. Each of the crude extracts (MeOH and $CH_2Cl_2$) of Limonium tetragonum showed a significant inhibitory effect on the growth of human cancer cells. The combined crude extracts of MeOH and $CH_2Cl_2$ were partitioned between $CH_2Cl_2$ and water. The organic layer was further partitioned between 85% aq. MeOH and n-hexane, and then the aqueous layer was fractionated with n-BuOH and $H_2O$, successively. Growth inhibition effects of crude extracts and their solvent fractions from Limonium tetragonum increased in a dose-dependent manner. Among them, 85% aq. MeOH, n-hexane and n-BuOH fractions revealed very good inhibition effects on the growth of human cancer cells. These results suggest that we can isolate active compounds from Limonium tetragonum to show much more strong anticancer activity.

• Journal of Nutrition and Health
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• v.43 no.2
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• pp.105-113
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• 2010

Ixeris dentata var. albiflora Nakai, a herbal plant, is often used to make a strong stomach as an antiphlogistic used when dyspepsia and to improve appetite in Korea and China. And also it is used for adult diseases such as diabetes and liver diseases as Korean traditional medicine. In this study, the composition and DPPH radical scavenging activities of the root of Ixeris dentata var. albiflora Nakai and its effects on cell viability on vero and chang cells were investigated. Moisture, crude ash, crude protein and crude lipid were 79.14, 2.49, 8.28 and 2.56 g/100 g respectively. The highest mineral content was K. The major free sugars were glucose, fructose and sucrose. Major fatty acid are linoleic acid, palmic acid and linolenic acid. Major amino acids were glutamic acid, arginine and aspartic acid and the total contents of amino acids were 28.12 mg/g. The methanol extracts were further fractionated with n-hexane, chloroform, ethylacetate, butanol and water to get an active fraction. In addition, cell viabilities in each fraction were determined. Methanol extract, butanol, and aqueous fraction showed strong survival rates in vero cell and chang cell viability test, and hexane, chloroform, and ethylacetate fraction were examined for toxin in a cell. The root of Ixeris dentata var. albiflora Nakai had scavenging activities against DPPH radicals in a dose-dependent assay. Ethylacetate fraction's SC50 was $6.8\{\mu}g/mL$, very strong DPPH radical scavenging activities, but water fraction did not show any activity.

• Journal of Life Science
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• v.27 no.9
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• pp.1064-1069
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• 2017

This study compared the cytotoxic effect of extracts from four different sea bream species (Pagrus major, Acanthopagus schlegeli, Oplegnathus fasciatus, and Girella punctata) in human cancer cell lines. Cytotoxic activity against the growth of human gastric adenocarcinoma (AGS) and HT-29 human colon cancer cell lines was determined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Treatment with acetone/methylene chloride (A+M) and methanol (MeOH) extracts from the four sea bream species dose-dependently increased cytotoxicity against the growth of AGS and HT-29 cancer cells (p < 0.05). As shown by a cell viability assay, treatment with A+M and MeOH extracts from red sea bream (P. major) had the highest cytotoxic effect (p < 0.05) among the sea bream species. The IC50 values of an 85% aqueous methanol (85% aq. MeOH) fraction from red sea bream (P. major) against AGS and HT-29 cancer cells was 0.33 and 1.58 mg/ml, respectively, suggesting that the 85% aq. MeOH fraction had the highest cytotoxic effect among the fractions (p < 0.05). Our results demonstrate that four different sea bream species exhibited cytotoxic activity, as well as high-quality amino acids and fatty acids. Among the sea bream species, red sea bream (P. major) showed the greatest cytotoxic effect. The results could be used to improve nutrition information available to consumers.

• Korean Journal of Soil Science and Fertilizer
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• v.31 no.4
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• pp.392-399
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• 1998

Nitrogen fertilizers such as urea are readily hydrolyzed in soils to produce ammonium ions which pass through nitrification and denitrification processes. These serial processes have drawn attention due to nitrogen losses, eutrophication, blue baby syndrome, and ozone depletion problems. The purpose of this study was to test the inhibitory effects of hot-water extract and organic solvent fractions of Artemisia asiatica leaves on soil urea hydrolysis and nitrification. In addition, the effects of organic solvent fractions on urease activity and ureolytic bacterial population were also investigated. First, hot-water extract of Artemisia asiatica leaves inhibited soil nitrification substantially with a marginal stimulatory effect on soil urea hydrolysis. Soils treated with hot-water extract of Artemisia asiatica leaves showed significant decreases in the accumulation of soil $NO_3-N$ (~68% decrease) compared with the control soil without the treatment of hot-water extract. In contrast, $CHCl_3$/MeOH fraction and basic aqueous layer of Artemisia asiatica leaves inhibited soil urea hydrolysis very strongly, causing 5.8 and 4.3-fold higher accumulation in amounts of remaining urea-N compared with the non-treated soil. Meanwhile, non of the organic solvent fractions showed any significant effects on soil nitrification inhibition. The inhibition of ureolytic bacterial activity by $CHCl_3$/MeOH fraction and aqueous basic layer of Artemisia asiatica leaves without any effects on urease activity itself led us to conclude that the inhibitions of soil urea hydrolysis were caused by the antagonistic effects on ureolytic bacterial activity.

• Microbiology and Biotechnology Letters
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• v.37 no.2
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• pp.118-124
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• 2009

Esterase EM2L8 gene isolated from deep sea sediment was expressed in Escherichia coli BL21 (DE3) and the esterase activity of the cell-free extract was assayed using p-nitrophenyl butyrate-spectrophotometric method. Its optimum temperature was $40-45^{\circ}C$ and 45% activity of the maximum activity was retained at $15^{\circ}C$. The activation energy at $15-45^{\circ}C$ was calculated to be 4.9 kcal/mol showing that esterase EM2L8 was a typical cold-adapted enzyme. Enzyme activity was maintained for 6 h and 4 weeks at $30^{\circ}C$ and $4^{\circ}C$, respectively. When each ethanol, methanol, and acetone was added to the reaction mixture to 15% concentration, enzyme activity was maintained. In the case of DMSO, enzyme activity was kept up to 40% concentration. (S)-4-Chloro-3-hydroxy butyric acid is a chiral intermediate for the synthesis of Atorvastatin, a hyperlipemia drug. When esterase EM2L8 (40 U) was added to buffer solution (1.2 mL, pH 9.0) containing ethyl-(R,S)-4-chloro-3-hydroxybutyrate (38 mM), it was hydrolyzed into 4-chloro-3-hydroxy butyric acid with a rate of $6.8\;{\mu}mole/h$. The enzyme hydrolyzed (S)-substrate more rapidly than (R)-substrate. When conversion yield was 80%, e.e.s value was 40%. When DMSO was added, hydrolysis rate increased to $10.4\;{\mu}mole/h$. The plots of conversion yield vs e.e.s in the presence or absence of DMSO were almost same, implying that the reaction enantioselectivity was not changed by the addition of DMSO. Taken together, esterase EM2L8 had high activity and stability at low temperatures as well as in various organic solvents/aqueous solutions. These properties suggested that it could be used as a biocatalyst in the synthesis of useful pharmaceuticals.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.3
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• pp.480-486
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• 2004

Despite many therapeutic advances in the understanding of the processes in carcinogenesis, overall mortality statistics are unlikely to change until there is reorientation of the concepts for the use of natural products as new anticarcinogenic agents. In this study, we investigated the anticarcinogenic activity, antioxidant and DPPH scavenging activity of Sargassum fulvellum (SF). SF was extracted with methanol, which was further fractionated into five different types: hexane (SFMH), ethylether (SFMEE), ethyl acetate (SFMEA), butanol (SFMB) and aqueous (SFMA) partition layers. We determined the cytotoxic effect of these layers on human cancer cells by MTT assay. Among various partition layers of SF, at starting concentration of 100 $\mu\textrm{g}$/mL, SFMEE showed very high cytotoxicity which were 92, 90 and 84% and kept high throughout 5 concentration levels sparsed by 100 $\mu\textrm{g}$/mL against all three human cancer cell lines: HepG2, HT-29 and HeLa. SFMEA showed a low cytotoxicity at the beginning concentration level, but as the concentration became denser, growth inhibition effect of cancer cell lines started to increase and at 500 $\mu\textrm{g}$/mL, it hit the highest, which were 91, 96 and 98% against the same three cell lines as above. We observed QR induced effect in all fraction layers of SF. SFMEE showed similar tendensy of QR induced effect as did against cytotoxicity. The QR induced effect of SFMEE on HepG2 cells at 25 $\mu\textrm{g}$/mL concentration indicated 3 times higher than the control value of 1.0 and SFMH tended to be concentration-dependent on HepG2 cells. At 100 $\mu\textrm{g}$/mL, the QR induced effects resulted a ratio, which was 2.5 times higher than the control value. In search for antioxidation effects of SF extract and partition layer, the reducing activity on the 1, 1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging potential was sequentially screened. The SFM has similar antioxidant activity as to BHT and vitamin C groups.

• Horticultural Science & Technology
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• v.29 no.4
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• pp.358-365
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• 2011

Lespedeza cuneata G. Don is a plant commonly grown in Asian countries, which has been widely used as an oriental medicinal herb to treat diabetes, diarrhea and various other inflammatory diseases. The phenolics of dry leaves of L. cuneata G. Don were extracted by using 80% (v/v) aqueous methanol in assistance with homogenization and sonification. The phenolic extract and its five different fractions (n-hexane, chloroform, ethyl acetate, n-butanol, and water) were used to evaluate the levels of total phenolics, total flavonoids, and antioxidant capacity as well as the inhibitory effect of tyrosinase activity. Ethyl acetate fraction (1 g) had the highest levels of total phenolics at 240.8 mg gallic acid equivalents (GAE), total flavonoids as 90.4 mg catechin equivalents (CE) as well as antioxidant capacity at 523.4 mg vitamin C equivalents (VCE) on ABTS assay and 329.5 mg VCE on DPPH assay among fractions. One g of water fraction contained total phenolics at 133.1 mg GAE, total flavonoids at 34.5 mg CE, and antioxidant capacity at 333.4 mg VCE for ABTS assay and 313.2 mg VCE for DPPH assay. Inhibition of tyrosinase activity of water fraction at 300 ${\mu}g{\cdot}mL^{-1}$ was at 47.2% and 21.1% for L-tyrosine and L-DOPA as its substrate, respectively. On the other hand, ethyl acetate fraction at 300 ${\mu}g{\cdot}mL^{-1}$ showed tyrosinase inhibition of 10.2% for L-tyrosine and 11.9% for L-DOPA. These results suggested that the phenolics from dry leaves of L. cuneata G. Don may be utilized as a potent source of antioxidants and skin whitening agents.

• Journal of Applied Biological Chemistry
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• v.60 no.1
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• pp.5-11
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• 2017

18 flowers of Compositae family were collected and extracted in aqueous methanol (MeOH). The concentrated extract was partitioned into n-hexane, ethyl acetate (EtOAc), n-BuOH, and water fractions. The extract and fractions were evaluated for total phenolics, total flavonoids, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, and tyrosinase inhibition activity. n-Hexane and EtOAc fractions of Aster yomena, n-hexane fraction of Cosmos bipinnatus White, n-hexane and EtOAc fractions of C. bipinnatus Pink showed high total phenolics. And EtOAc fractions of A. yomena, C. bipinnatus White, C. bipinnatus Red, C. morifolium Froggy, and C. morifolium Himaya exhibited high total flavonoids. EtOAc fractions of A. yomena, C. bipinnatus White, C. bipinnatus Pink, C. morifolium Yellowmable, and MeOH extract of C. morifolium Rosa significantly scavenged DPPH radical. EtOAc fractions of C. chinensis, C. bipinnatus White, C. bipinnatus Red, C. morifolium Himaya, and C. morifolium Hongsim highly inhibited the tyrosinase activity. A. yomena, C. bipinnatus White, C. bipinnatus Pink, C. bipinnatus Red and C. morifolium Himaya are evaluated as good source for whitening cosmetics materials.